Thesis

Chapter-2 Literature Review
1. INTRODUCTION
Eye is most interesting organ due to its drug disposition characteristics. Generally,
topical application of drugs is the method of choice under most circumstances because of its
convenience and safety for ophthalmic chemotherapy.
!"
# significant challenge to the formulator is to circumvent bypass" the protective
barriers of the eye without causing permanent tissue damage. $evelopment of newer, more
sensitive diagnostic techni%ues and novel therapeutic agents continue to provide ocular
delivery systems with high therapeutic efficacy. Conventional ophthalmic formulations li&e
solution, suspension, and ointment have many disadvantages which result into poor
bioavailability of drug in the ocular cavity. 'he specific aim of designing a therapeutic system
is to achieve an optimal concentration of a drug at the active site for the appropriate duration.
2"

(cular disposition and elimination of a therapeutic agent is dependent upon its
physicochemical properties as well as the relevant ocular anatomy and physiology. #
successful design of a drug delivery system, therefore, re%uires an integrated &nowledge of
the drug molecule and the constraints offered by the ocular route of administration.)"'he
various approaches that have been attempted to increase the bioavailability and the duration
of the therapeutic action of ocular drugs can be divided into two categories.
'he first one is based on the use of sustained drug delivery systems, which provide
the controlled and continuous delivery of ophthalmic drugs. 'he second involves ma*imi+ing
corneal drug absorption and minimi+ing precorneal drug loss.)" ,deal ophthalmic drug
delivery must be able to sustain the drug release and to remain in the vicinity of front of the
eye for prolong period of time. Conse%uently it is imperative to optimi+e ophthalmic drug
delivery- one of the way to do so is by addition of polymers of various grades, development
of in situ gel or colloidal suspension or using erodible or non erodible insert to prolong the pre
corneal drug retention.."
1.1. IN SITU FORMING GELS FOR OPTHALMIC DRUG DELIVERY:
Recently, controlled and sustained drug delivery has become the standard in modern
pharmaceutical design and an intensive research has been under ta&en in achieving much
better drug product effective ness reliability and safety. ,n this regard many polymers are very
useful which undergo reversible sol to gel phase transition in response to physiological
stimuli./" ,n situ gels are conveniently dropped as a solution into the con0unctival sac, where
they underego a transition into a gel with its favourable residence time. 'he sol-gel transition
occurs as a result of a chemical1 physical change induced by physiological environment. 'his
Mahathi College of Pharmacy, Madanapalle Page 1
type of gel combines the advantage of a solution being patient convenient with the favourable
residence time of a gel for enhancing the ocular bioavailability 2".
'he sol-gel transition can be induced by a shift in the p3 as for cellulose acetate
phthalate, a shift in temperature as for the thermogelling 4olo*amer !55 or by presence of
cations as for deacetylated gellan gum and alginates. 'hus, the in situ gelling systems for
ophthalmic use can be classified as p3 sensitive, temperature sensitive and ion-activated
systems. 'he rate of gel formation in situ, is important since when dropped in the eye, before
a strong gel is formed, a solution or a wea& gel is prone to elimination by the fluid mechanics
of the eye 5".
'he ion activated in situ gelling system can be formulated using sodium alginate, the
sodium salt of alginic acid, as a natural hydrophilic polysaccharide containing two types of
monomers, 6-$-mannuronic acid 7" and *-L-guluronic acid G" which forms a gel in the
cul-de-sac due to the presence of divalent calcium ions in the lacrimal fluid 8". 'hus with the
use of these in situ gelling systems, residence time of the drug in the eye is increased.
Continuous delivery of drugs in a controlled manner to the anterior chamber of the eye will
eliminate the re%uirement for fre%uent drug administration, causing better patient compliance
and will result in e*tended duration of action, hence lower amount of total dose re%uired,
which in turn will minimi+e the local and1or systemic side effects !9".
1.2. THE ANATOMY OF THE EYE:
'he human eye, elegant in its detail and design, represents a gateway to the process
we call vision. 'he eyeball is spherical in shape and about ! inch across. ,t houses many
structures that wor& together to facilitate sight. 'he human eye is comprised of layers and
internal structures, each of which performs distinct functions. 'he detailed description of each
eye part is given below.
Fig. No. 1: Anatom o! ""
A. S#$"%a
'he sclera white portion of the eye" is the tough white sheath that forms the outer-
layer of the ball. ,t is a firm fibrous membrane that maintains the shape of the eye as an
appro*imately globe shape. ,t is much thic&er towards the bac&1posterior aspect of the eye
than towards the front1anterior of the eye. !!"
&. Con'(n#ti)a
'he con0unctiva is a thin transparent mucous epithelial barrier, lines the inside of the
eyelids, and covers the anterior one-third of the eyeball. 'he respective portion of con0unctiva
is referred to as the palpebral and bulbar con0unctiva. 'he con0unctiva is composed of two
layers: an outer epithelium and its underlying stroma substantia propria". 'he e*posed
surface of the eye includes con0unctiva and cornea and is covered with the tear film. 'he
con0unctiva contributes to the formation of the tear film by way of secreting substantial
electrolytes, fluid, and mucins.
C. Co%n"a
'he cornea is a strong clear bulge located at the front of the eye. ;urface of the adult
cornea has a radius of appro*imately 5mm. ,t has an important optical function as it refracts
light entering the eye which then passes through the pupil and onto the lens which then
focuses the light onto the retina". 'he cornea, a non-vascular structure does not contain any
blood vessels" gets the necessary nutrients from the capillaries that terminate in loops at its
circumference. ,t is supplied by many nerves derived from the ciliary nerves. 'hese enter the
laminated tissue of the cornea. ,t is therefore e*tremely sensitive.
D. A*("o(+ ,(mo%
Fig. No. 2: Pat,-a o! A*("o(+ H(mo%
'he a%ueous humor is a 0elly-li&e substance located in the outer1front chamber of the
eye. ,t is a watery fluid that fills the <anterior chamber of the eye< which is located
immediately behind the cornea and in front of the lens. 'he a%ueous humor is very slightly
al&aline salt solution that includes tiny %uantities of sodium and chloride ions. ,t is
continuously produced, mainly by the ciliary processes, flows from the posterior chamber
through the pupil into the anterior chamber, and e*its via the trabecular route at the angle and
the uveoscleral route. ;chlemm=s canal canal of ;chlemm or the scleral venous sinus", is a
circular channel that collects a%ueous humour from the anterior chamber and delivers it into
the bloodstream via the anterior ciliary veins. ,t is located at the 0unction of the cornea and the
sclera. ,n human, the rate of a%ueous humor turnover is appro*imately !> - !.?> of the
anterior chamber volume per minute. 'he rate of a%ueous formation is appro*imately 2.?
@l1min. #%ueous humor consists of pressure dependent and pressure independent pathways.
'he pressure dependent outflow refers to the trabecular meshwor&-schlemmAs canal-venous
system, while pressure independent outflow refers to any non trabecular outflow and is called
as uveoscleral outflow. !2"
E. P(.i$
4upil generally appears to be the dar& <centre< of the eye, but can be more accurately
described as the circular aperture in the centre of the iris through which light passes into the
eye. 'he si+e of the pupil and therefore the amount of light that is admitted into the eye" is
regulated by the pupillary refle* also &nown as the <light refle*<".
F. I%i+
'he iris is a thin circular contractile curtain located in front of the lens but behind the
cornea. 'he iris is a diaphragm of variable si+e whose function is to ad0ust the si+e of the
pupil to regulate the amount of light admitted into the eye. ,t is the coloured part of the eye
shades may vary individually li&e blue, green, brown, ha+el, or grey".
G. Ci$ia% M(+#$"
'he ciliary muscle is a ring of striated smooth muscles in the eyeAs middle layer that
controls accommodation for viewing ob0ects at varying distances and regulates the flow of
a%ueous humour into schlemmAs canal. 'he muscle has parasympathetic and sympathetic
innervation. Contraction and rela*ation of the ciliary muscle alters the curvature of the lens.
'his process may be described simply as the balance e*isting at any time between two states:
Ciliary 7uscle rela*ed 'his enables the eye to focus on distant ob0ects" and Ciliary 7uscle
contracted 'his enables the eye to focus on near ob0ects".
H. L"n+
'he lens is a transparent structure enclosed in a thin transparent capsule. ,t is located
behind the pupil of the eye and encircled by the ciliary muscles. ,t helps to refract light
travelling through the eye which first refracted by the cornea". 'he lens focuses light into an
image on the retina. ,t is able to do this because the shape of the lens is changed according to
the distance from the eye of the ob0ects" the person is loo&ing at. 'his ad0ustment of shape of
the lens is called accommodation and is achieved by the contraction and rela*ation of the
ciliary muscles.
I. Vit%"o(+ H(mo(%
'he vitreous humour also &nown as the vitreous body" is located in the large area
that occupies appro*imately 59> of each eye in the human body. 'he vitreous humour is a
perfectly transparent thin-0elly-li&e substance that fills the chamber behind the lens of the eye.
,t is an albuminous fluid enclosed in a delicate transparent membrane called the hyaloid
membrane.
/. R"tina
'he retina is located at the bac& of the human eye. 'he retina may bedescribed as the
<screen< on which an image is formed by light that has passed into the eye via the cornea,
a%ueous humour, pupil, lens, and finally the vitreous humour before reaching the retina. 'he
function of the retina is not 0ust to be the screen onto which an image may be formed but also
to collect the information contained in that image and transmit it to the brain in a suitable
form for use by the body. 'he retinal <screen< is therefore a light-sensitive structure lining
the interior ofthe eye. ,t contains photosensitive cells called rods and cones" and their
associated nerve fibers that convert the light they detect into nerve impulses that are then sent
onto the brain along the optic nerve.
0. Ma#($a
'he center of the retina is called the macula. 'he macula contains a high
concentration of photoreceptor cells which convert light into nerve signals. Because of the
high concentration of photoreceptors, we are able to see fine details such as newsprint with
the macula. #t the very center of the macula is the fovea, the site of our sharpest vision.
L. C,o%oi1
Fig. No. 2: Po+t"%io% )i"- o! ""
'he choroid layer is located behind the retina and absorbs unused radiation and
nourishes the outer portions of the retina. ,t is a thin, highly vascular i.e. it contains blood
vessels" membrane that is dar& brown in colour and contains a pigment that absorbs e*cess
light and so prevents blurred vision due to too much light on the retina". 'he choroid has one
of the highest blood flows in the body. 'he choroid is loosely attached to the inner surface of
the sclera by the lamina fusa.
M. O.ti# n"%)"
'he optic nerve a bundle of over ! million nerve fibers" is responsible for
transmitting nerve signals from the eye to the brain. 'hese nerve signals contain information
on an image for processing by the brain. 'he front surface of the optic nerve, which is visible
on the retina, is called the optic dis&.
1.2.1. A##"++o% o%gan+ o! t," "":
'he eye is protected by several structures.
Eyebrows
Eyelids and eyelashes
Lacrimal apparatus
Eyebrows protect the anterior aspect of eyeball from sweat, dust and foreign bodies.
'he eyelids have various layers of tissue including con0unctiva which protects the delicate
cornea and front of the eye. Chen eye drops are administered, they are placed in lower
con0unctival sac. 'he lacrimal glands secrete tears composed of water, mineral salts,
antibodies and lyso+yme, a bactericidal en+yme. $rainage of the eye drops through
nasolacrimal system into gastrointestinal tract begins immediately on instillation. 'his ta&es
place when either refle* tearing or the dosage form causes volume of fluid in peripheral tissue
to e*ceed the normal lacrimal volume of 2-!9 Dl. 'he e*cess fluid volume enters the superior
and inferior lacrimal puncta, moves down the canalicula into the lacrimal sac, and continues
into the gastrointestinal tract!)".
1.2. ROUTES OF OCULAR DRUG DELIVERY
'here are several possible routes of drug delivery into the ocular tissues. 'he
selection of the route of administration depends primarily on the target tissue.
1.2.1. To.i#a$ %o(t"
'ypically topical ocular drug administration is accomplished by eye drops, but they
have only a short contact time on the eye surface. 'he contact, and thereby duration of drug
action, can be prolonged by formulation design e.g.m gels, gelifying formulations, ointments,
and inserts".
Fig. No. 3: Di!!"%"nt Ro(t"+ !o% O#($a% D%(g D"$i)"%
1.2.2. S(4#on'(n#ti)a$ a1mini+t%ation
'raditionally subcon0unctival in0ections have been used to deliver drugs at increased
levels to the uvea. Currently this mode of drug delivery has gained new momentum for
various reasons. 'he progress in materials sciences and pharmaceutical formulation have
provided new e*citing possibilities to develop controlled release formulations to deliver drugs
to the posterior segment and to guide the healing process after surgery.
1.2.2. Int%a)it%"a$ a1mini+t%ation
$irect drug administration into the vitreous offers distinct advantage of more
straightforward access to the vitreous and retina. ,t should be noted- however that delivery
from the vitreous to the choroid is more complicated due to the hindrance by the R4E Retinal
4igment Epithelium" barrier. ;mall molecules are able to diffuse rapidly in the vitreous but
the mobility of large molecules, particularly positively charged, is restricted.
1.3. &ARRIERS FOR OCULAR DELIVERY:
1.3.1. D%(g $o++ !%om t," o#($a% +(%!a#"
#fter instillation, the flow of lacrimal fluid removes instilled compounds from the
surface of eye. Even though the lacrimal turnover rate is only about ! @l1min the e*cess
volume of the instilled fluid is flown to the nasolacrimal duct rapidly in a couple of minutes.
#nother source of non-productive drug removal is its systemic absorption instead of ocular
absorption. ;ystemic absorption may ta&e place either directly from the con0unctival sac via
local blood capillaries or after the solution flow to the nasal cavity.
1.3.2. La#%ima$ !$(i15"" 4a%%i"%+
Corneal epithelium limits drug absorption from the lacrimal fluid into the eye. 'he
corneal epithelial cells form tight 0unctions that limit the paracellular drug permeation.
'herefore, lipophilic drugs have typically at least an order of magnitude higher permeability
in the cornea than the hydrophilic drugs. ,n general, the con0unctiva is lea&ier epithelium than
the cornea and its surface area is also nearly 29 times greater than that of the cornea.
1.3.2. &$oo15o#($a% 4a%%i"%+
'he eye is protected from the *enobiotics in the blood stream by blood-ocular
barriers. 'hese barriers have two parts: blood-a%ueous barrier and blood-retina barrier. 'he
anterior blood-eye barrier is composed of the endothelial cells in the uveam 'he middle layer
of the eye beneath the the sclera. ,t consists of the iris, ciliary body, and choroid".
'his barrier prevents the access of plasma albumin into the a%ueous humor, and also
limits the access of hydrophilic drugs from plasma into the a%ueous humor. 'he posterior
barrier between blood stream and eye is comprised of retinal pigment epithelium R4E" and
the tight walls of retinal capillaries. Enli&e retinal capillaries the vasculature of the choroid
has e*tensive blood flow and lea&y walls. $rugs easily gain access to the choroidal
e*travascular space, but thereafterdistribution into the retina is limited by the R4E and retinal
endothelia.
Ta4$" 1: &a%%i"%+ !o% t," O#($a% 1"$i)"%
1.6. MECHANISM OF OCULAR DRUG A&SORPTION 7138
$rugs administered by instillation must penetrate the eye and do so primarily through
the cornea followed by the non-corneal routes. 'hese non-corneal routes involve drug
diffusion across the con0unctiva and sclera and appear to be particularly important for drugs
that are poorly absorbed across the cornea.
Con'(n#ti)a
Co%n"a S#$"%a
;urface area
!2./? F 2.!2 cm
2
!.9. F 9.!2 !/ G !2
'hic&ness
- 9.?2 mm 9.. -9.? mm
;tructural
composition
7ucus membrane
Epithelium
Hasculature
? layers
Epithelium
BowmanAs
membrane
;tomata
$escemetAs
membrane
Endothelium
Collagen
fibers
Cater
4roteoglycans
7onopolysaccharides
Elastic fibers
Iibroblast
Fig. No. 6: O#($a% D%(g A4+o%.tion
1.6.1. Co%n"a$ ."%m"ation
'he permeation of drugs across the corneal membrane occurs from the precorneal
space.
Fig. No. 9: Co%n"a$ M"m4%an" D".i#ting
1.9. Va%io(+ &a%%i"i%+ to 1%(g A4+o%.tion:
,n tears have a direct bearing on efficiency of drug absorption into the inner eye.'he
productive absorption of most ophthalmic drugs results from diffusional process across
corneal membrane.'he efficiency of absorption process is a function of rate and e*tent at
which the transport processes of eye. 'he flu* of any drug molecule across the biological
membrane depends on the physicochemical properties of the permeating molecule and its
interaction with the membrane. 'he e*tent to which the transport or absorption process occurs
is also function of physiological mechanism of precorneal fluid drainage or turnover. ,n
terms of transcorneal drug permeation, the cornea can be considered to consist of three
primary layers epithelium, stroma and endothelium".
'he epithelium and endothelium contain on the order of a !99 fold greater amount of
lipid material than the stroma. Conse%uently, depending on the physicochemical properties of
a diffusing drug, the resistance offered by the individual layers varies greatly. Epithelium,
being lipodal, represents a diffusional barrier offering high resistance to ionic or other
a%ueous soluble or polar species. ,n contrast, compounds with relatively low polarity
encounter a greater diffusional resistance in the hydrophilic stroma layer. 'his fre%uently
cited concept of drug permeation across the corneal membrane ism referred to as Jdifferential
solubility conceptK.
1.9.1. Non5#o%n"a$ ."%m"ation
4rimary mechanism of drug permeation is the sclera is li&ely to be diffusionacross the
intercellular a%ueous media in the case of structurally similar corneal stroma. 'herefore the
possibility of partitioning mechanism cannot be eliminated. #lthough li&e cornea, the
con0unctiva is composed of an epithelial layer covering an underlying stroma, the
con0unctival epithelium offers substantially less resistance than does the corneal epithelium.
1.9.2. Va%io(+ !a#to%+ %"+.on+i4$" !o% 1i+.o+ition o! o#($a% 1%(g+: 7138
Bioavailability of drugs administered to the eye is an important consideration. 'here
are physiological factors, which can affect a drugAs bioavailability including protein binding,
drug metabolism and lachrymal drainage.
4rotein bound drugs are incapable of penetrating the corneal epithelium due to the
si+e of the protein drug comple*. Because of the brief time in which an ophthalmic solution
may remain present in the eye due to lachrymal drainage", protein binding of a drug
substance could %uic&ly negate its therapeutic value by rendering it unavailable for
absorption. (ne of the ma0or problems encountered with conventional ophthalmic solutions
is the rapid and e*tensive elimination of drugs from the precorneal lachrymal fluid. ,t must be
noted that this high drainage rate is due to the tendency of the eye to maintain its residence
volume at 2G!9 @l permanently, whereas volumes topically instilled range from 29G?9 @L. ,n
fact it has been demonstrated in vivo that 89> of the dose was cleared within 2 min for an
instilled volume of ?9 @L and, within . min for an instilled volume of !9 @l. Conse%uently,
the ocular residence time of conventional solutions is limited to a few minutes, and the overall
absorption of a topically applied drug is limited to !G!9>. #s in the case with other
biological fluids, tears contain en+ymes such as lyso+ymes" capable of the metabolic
degradation of the drug substance.
,n addition to the physiological factors affecting ocular bioavailability, other factors
as the physicochemical properties of the drug substance, and product formulation are
important. Because the cornea is a membrane-barrier containing both hydrophilic and
lipophilic layers, it is permeated most effectively by drug substances having both hydrophilic
and lipophilic characteristics. ,t is advantageous for corneal penetration to ad0ust the p3 of the
solution to increase the proportion of unioni+ed drug in the instilled dose. $rugs, which are
highly water insoluble, do not readily permeate the cornea.
1.9.2. Na+o$a#%ma$ 1%ainag" ++t"m: 7168
'he nasolachrymal drainage system consists of three parts: the secretory system, the
distributive system and the e*cretory system. 'he secretory system consists of basic secretors
that are stimulated by blin&ing and temperature change due to tear evaporation and refle*
secretors that have an efferent parasympathetic nerve supply and secrete in response to
physical or emotional stimulation.
Fig. No. : Na+ao$a#,%ma$ D%ainag" A..a%at(+
'he distributive system consists of the eyelids and the tear meniscus around the lid
edges of the open eye, which spread tears over the ocular surface by blin&ing, thus preventing
dry areas from developing. 'he e*cretory part of the nasolachrymal drainage system consists
of: the lachrymal puncta, the superior, inferior and common canaliculi- the lachrymal sac and
the nasolachrymal duct. ,n humans, the two puncta are the openings of the lachrymal
canaliculi and are situated on an elevated area &nown as the lachrymal papilla. ,t is thought
that tears are largely absorbed by the mucous membrane that lines the ducts and the lachrymal
sac only a small amount reaches the nasal passage.
1.:. Int"%"+t+ o! no)"$ o.,t,a$mi# 1%(g 1"$i)"%:
(phthalmic drug delivery is one of the most interesting and challengingendeavors
facing the pharmaceutical scientist. !." 'he landscape of ophthalmic drug delivery is highly
competitive and rapidly evolving. Lew classes of pharmaceuticals and biologics are fueling
the demand for novel drug delivery.
'he main aim of pharmacotherapeutics is the attainment of effective drug
concentration at the site of action for the sufficient period of time to elicit a response. 'he
challenge is to provide a system with improved ocular drug bioavailability and prolonged
duration of activity, but still with a minimum ris& of ocular complications. # ma0or problem
of ophthalmic drug delivery is not the lac& of efficient drugs but the attainment of their
optimal concentration at the site of their optimal concentration at the site of action. !?"
'he emergence of new and innovative means for improving therapeutic efficacy
suggests that a greater choice of dosage forms will be provided to physicians and patients in
the ne*t decade. 7ost of the formulation efforts aim at ma*imi+ing ocular drug absorption
through prolongation of the drug residence time in the cornea and con0unctival sac, as well as
to slow drug release from the delivery system and minimi+e precorneal drug loss. Harious
ophthalmic formulations and their residence time period in the ocular cavity are given below.
!/"
Fig. No. ; D(%ation o! a#tion o! o#($a% 1%(g 1"$i)"% ++t"m+
1.;. O.,t,a$mi# 1%(g !o%m($ation+:
(phthalmic drugs are formulated to bring the active drugs in contact with the eye
surface to allow for absorption. E*tension of corneal contact time may result in increased
drug penetration M higher intraocular drug delivery. ,n addition to the active drug, ophthalmic
formulations should contain other ingredients to control various characteristics of the
formulation, such as the buffering and p3, osmolality M tonicity, viscosity M antimicrobial
preservatives. #lthough these ingredients are listed inactive, they can affect permeability of
drug across the ocular surface barriers M alter the therapeutic effectiveness of the drug.
1.<. EYE INFECTIONS:
Eyes can get infections from bacteria, fungi or viruses. Eye infections can occur in
parts of the eye and can affect 0ust one eye or both. Common eye infections are
Con0unctivitis, Corneal ulcers M Endophthalmitis.
A8 Con'(n#ti)iti+:
Con0unctivitis is swelling inflammation" or infection of the membrane lining the
eyelids con0unctiva".,t is characteri+ed by cellular infiltration and e*udation. ;taphylococcus
aureus is the most common cause of bacterial con0unctivitis and blepharo-con0unctivitis.
7any other organisms li&e 3aemophilus influen+ae, ;treptococcus pneumoniae also cause
con0unctivitis. Con0unctivitis can be classified as !" ,nfective G #cute, ;ubacute M
Chronic
2" #llergic con0unctivitis.
&8 Co%n"a$ ($#"%+= 0"%atiti+:
,nflammation of cornea Neratitis" is characteri+ed by corneal oedema, cellular
infiltration M ciliary congestion. Being the most anterior part of eyeball, cornea is e*posed to
atmosphere M hence prone to get infected easily. Bacterial corneal ulcers are the most
commonly caused by virulent organism. Common bacteria associated with corneal ulceration
are ;taphylococcus aureus, 4seudomonas pyocyanea, E.coli and 4roteus etc.
C8 En1o.,t,a$miti+:
,t is severe form of intraocular inflammation purulent uveitis" involving ocular
cavities M inner coats of eyeball. Causative organisms include ;treptococci, E.coli,
4seudomonas, etc.
#ccordingly, the armamentarium of available antimicrobials used in the prevention
and treatment of these infections includes antivirals, antifungals, and antibacterials. Common
topical antibacterials used in the treatment of ocular infectious diseases include sulfonamides,
aminoglycosides, polymy*in-based combinations, and !$(o%o*(ino$on"+.
'he fluoro%uinolones represent an e*panding class of broad-spectrum antibacterials,
which cover a host of Gram-negative and anaerobic species responsible for ocular infections.
'hese antibacterials have gained popularity in them ophthalmology field since they have been
shown to be e%uivalent to combination therapy in the treatment of many ocular infections.
Iluoro%uinolones are also effective against a variety of Gram-positive organisms, including
;treptococcal and ;taphylococcal species- however, resistance is emerging among some of
these organisms. 'he classification and mechanism of action of fluoro%uinolones are given
below
Ta4$" 2: Common$ U+"1 F$(o%o*(ino$on"+ in O.,t,a$mi# D"$i)"%
Anti 4ioti#
g"n"%ation
E>am.$" A#ti)it
!
st
GELE#R',(L
Lalidi*ic acid

3ave limited activity against
gram negative M gram positive
organism
2
nd
GELER#',(L
(*olinic acid
Cino*acin
4ipemic acid
,mprovement in gram negative
coverage including
#ntipseudomonal activity.
;hows limited activity against
Gram positive organism.
)
rd
GELER#',(L
Lorflo*acin
Ciproflo*acin
Leavoflo*acin
(flo*acin
3aving antipseudomonal
activity against gram negative
bacilli
.
th
GELER#',(L
Ciproflo*acin
7o*iflo*acin
Gatiflo*acin
3aving dual mechanism of action
in gram positive bacteria in
addition reducing efflu* from the
bacterial cell.
,mproved spectrum of #ctivity.
1.1?. MANAGEMENT OF OCULAR INFECTIONS
(cular infections, both superficial and deep such as con0unctivitis, corneal ulcers and
endophthalmitis are caused by diverse group of bacteria, viral and fungal pathogens.
#ccordingly the armamentarium of available antimicrobials used in the prevention and
treatment of these infections includes antivirals, antifungals and antibacterials. Common
topical antibacterials used in treatment of ocular infectious diseases include sulfonamides,
aminoglycosides, polymy*in-based combinations and fluoro%uinolones. 'hese
fluoro%uinolones are indicated for severe bacterial &eratitis, endophthalmitis, blepharo-
con0unctivitis, corneal ulcers, chronic post-filtration hypotony etc. 'he fluoro%uinolones
represent an e*panding class of broad spectrum antibacterials which cover a host of Gram
negative and anaerobic species responsible for ocular infections. 'hese antibacterials have
gained popularity in the ophthalmology field since they have been shown to be e%uivalent to
combination therapy in treatment of many ocular infections. Iluoro%uinolones are also
effective against a variety of Gram positive organisms including ;treptococcal and
;taphylococcal species. !5"
Iluoro%uinolones offer all the attributes of an ideal antimicrobial agent including
broad antimicrobial spectrum, good tissue penetration and bioavailability, high rate of
clearance, chemical and biological stability, low degree of to*icity, high binding affinity for
melanin, better patient compliance, convenient dosage forms and dosing schedule and
relatively low incidence of drug interactions.
1.11. MECHANISM OF ACTION
Iluoro%uinolones act by inhibiting two en+ymes involved in bacterial $L# synthesis,
both of which are $L# topoisomerases that human cells lac& and that are essential for
bacterial $L# replication, thereby enabling these agents to be both specific and bactericidal.
$L# topoisomerases are responsible for separating the strands of duple* bacterial $L#,
inserting another strand of $L# through the brea&, and then resealing the originally separated
strands.
$L# gyrase introduces negative superhelical twists in the bacterial $L# doubleheli*
ahead of the replication for&, thereby cataly+ing the separation of daughter chromosomes.
'his activity is essential for initiation of $L# replication and allows for binding of initiation
proteins. 'opoisomerase ,H is responsible for decatenation that is, removing the interlin&ing
of daughter chromosomes thereby allowing segregation into two daughter cells at the end of a
round of replication. Iluoro%uinolones interact with the en+yme-bound $L# comple* i.e.,
$L# gyrase with bacterial $L# or topoisomerase ,H with bacterial $L#" to create
conformational changes that result in the inhibition of normal en+yme activity.
#s a result, the new drugG en+ymeG$L# comple* bloc&s progression of the
replication for&, thereby inhibiting normal bacterial $L# synthesis and ultimately resulting in
rapid bacterial cell death.!8" (lder fluoro%uinolones e*hibit a relatively consistent pattern
with respect to specificity of en+yme inhibition in different types of bacteria. 'he newer
fourth generation fluoro%uinolones li&e mo*iflo*acin, gatiflo*acin have a dual-binding
mechanism of action, inhibiting both $L# gyrase and topoisomerase ,H, in Grampositive
species. 29"
1.12. Po$m"%i# 1%(g 1"$i)"%
3ydrogels are one of the upcoming classes of polymer-based controlled release drug
delivery systems.2!" 4olymeric drug delivery systems have been e*tensively studied in order
to solve the potential problems associated with drugs or bioactive molecules including
to*icity, site dependence, low effectiveness, poor solubility, short half life, rapid degeneration
and rapid clearance from the body.
Considering various properties such as fle*ibility, structure, biocompatibility, and
hydrophilicity, three dimensional matrices, hydrogels, are being e*tensively used as drug
delivery carriers. 22"
1.12.1. A1)antag"+ o! .o$m"%i# 1%(g 1"$i)"%
Reduce to*ic effects on the healthy tissue and reach sites that are conventionally
,naccessible due to the presence of various barriers 8 by targeted drug delivery.
,ncrease the half-life of drugs, preventing their rapid degradation, and reduce the rate of
elimination, thus maintaining drug concentration within a therapeutically effective
window.
Reduce the amount of drug re%uired to achieve therapeutic efficacy.
Cut down the number of repeated invasive dosage re%uired for certain conditions and thus
helps to improve patientAs compliance and offers better living.
2. LITERATURE REVIE@
/aa%a' 0(ma%.0 "t a$ 72?1?8.A ;tudied ,n situ gels containing itracona+ole based on
temperature induced systems for the treatment of oral candidiasis. . 'he system using
pola*amer !9> to!?>w1w" along with carbopol 8). 9.9! to 9.. > w1w" and ,tracona+ole
!>w1w". 'he formulations were evaluated for physiochemical parameter, gelation,
temperature, viscosity, gel strength, content uniformity mucoadhesive force, I',R and $;C.
/aa%a' 0(ma%.0 "t a$ 72?1?8.A ,nvestigated the in situ gels system were formulated using
different concentration of gellan gum 9.! to 9.)", sodium alginate 9.! to 9.?" and carbopol
8). 9.! to 9.2" along with cicloprio* olamine !>".the viscosity was found to be in the range
to 8? Cps for the solution, whereas for the gels, it was up to 22999 cps. 'he optimi+ed
formulations were evaluated for gelling capacity, viscosity, gel strength, bioadhesive force,
spreadability, drug release, and I',R and $;C. ;tudies were carried out.
In1%a'""t D. Gon'a%i1 "t a$ 72??<8.A ;tudied ophthalmic gel formulations of flucona+ole.
,ntraocular delivery of topically applied drugs such as flucona+ole is hampered by elimination
of the solution due to tear turnover, so an in situ gelling thermoreversible mucoadhesive gel
was formulated. 'hermoreversible mucoadhesive gels were prepared using the cold method
along with polo*amer .92 and different mucoadhesive polymers such as hydro*y ethyl
cellulose 3EC", hydro*y propyl methyl cellulose 347C" N.7, and polyvinyl pyrrolidone
4H4" N)9. Gels were evaluated for physical parameters li&e appearance, gelation
temperature, p3, spreadability, drug content, gel strength, bioadhesion, and in vitro
permeation. # modified device modified N-C diffusion cell with a sheep=s eye corneal
membrane as a diffusion membrane" was used for evaluation of drug permeation through a
sheep=s corneal membrane. 'he formulated gels were transparent, uniform in consistency, and
had spreadability with a p3 range of /.5 to 2.). ;atisfactory bioadhesion on the sheep=s
corneal surface and good gel strength were also observed. $iffusion studies have shown that a
matri* is the best-fit model. #s the concentration of mucoadhesive agent increases, the rate of
permeation decreases. 'he order of drug permeation through the membrane was 3EC O 4H4
N)9 O 347C N.7. 'his study found that a thermoreversible polymer and mucoadhesive
polymers can be effectively used to prolong residence time.
B(Yang Li( "t a$ 72??<8.A 4repared 4olyethylene glycol"-polyP-caprolactone"-
polyethylene glycol" 4ECE" hydrogel was biocompatible and biodegradable despite of
temporary elevated intraocular pressure and slight corneal endothelial damage at specific
concentration.
Sa(%a4, G(.ta "t a$ 72??<".A $eveloped an ophthalmic delivery system of the fors&olin,
based on the concept of temperature activated in situ gelation. 4olo*amer .92 4luronic I-
!22" is a bloc& copolymer made of poly o*y ethylene" and poly o*y propylene". 'he sol-gel
transition is induced by an increase in temperature- however, it depends on the concentration
of the polymer and presence of other additives. 'he developed formulations were
therapeutically efficacious on albino Lew Qealand rabbit model"- reducing intra ocular
pressure ,(4" for !2 hours and showed sol-gel phase transition gelling" temperature of 22RC
and sustained drug release 22>F9.9?/> in vitro behavior over a period of . h.
Rai1a S. A$50a++a+ "t a$ 72??<8.A ;tudied was controlled release ophthalmic delivery
systems for ciproflo*acin ocular in situ gel carriers that undergo sol-to-gel transition upon
change in p3 or in the presence of cations in an attempt to prolong the effect of ciproflo*acin
and improve its ocular bioavailability. Carbopol and alginates polymers were used to confer
gelation properties to the formulations. 3ydro*ypropyl methylcellulose and methylcellulose
were combined with carbopol to increase the viscosity of the gels and to reduce the
concentration of the incorporated carbopol.
G(.ta H "t a$ 72??<8.A ,nvestigated Chitosan and 347C was characteri+ed for various in
vitro parameters. Ior scintigraphy study, the drug timolol maleate was radiolabeled 88m" 'c
by direct labeling method using ;nCl
2
.23
2
( as reducing agent. ,n vitro stability of the
radiolabeled drug 88m" 'c-timolol maleate comple*" was chec&ed and it was found to be
stable for up to 2. h. ,t was evident from the scintigraphic images and the time-activity curve
plotted from the data that the plain drug solution cleared very rapidly from the corneal region
and reached into systemic circulation via nasolachrymal drainage system, as significant
activity was recorded in &idney and bladder after 2 h of ocular administration. (cular
irritation of the developed formulation was also chec&ed by hen=s egg chorioallantoic
membrane test and formulation was found to be practically nonirritant.
&"%t o. Hag$(n1 "t a$ 72??<8.A ,nvestigated combination of Carbopol C" and methyl
cellulose 7C", was carried out at two different p3 ..9 and 2.." and temperatures 2? and
)2RC" by rotational cone and plate viscometry. 'he shear stress S" vs. shear rate $" flow
curves of the a%ueous polymer solutions indicated a pseudoplastic behavior, with a yield
point. #n increase in p3 from ..9 to 2.., or temperature from 2? to )2RC, resulted in an
increase in viscosity T", S, and yield point, the magnitude of changes being highest when
both the parameters were altered simultaneously.
Ha%i+, Nai% Mata.a1 "t a$ 72??<8.A Gel formulation of flucona+ole with mucoadhesive
properties is promising for prolonging bucal residence time and thereby better therapeutic
effects. ,n addition, they provide intimate contact between a dosage form and the absorbing
tissue, which may result in high drug concentration in local area. 'he in situ formulation will
have better patient acceptability since the formulation will be applied in the form of sols,
which upon contact will form the corresponding gels causing less irritation or pain.
/o+"., /ag(%5G%o1Cin+Dia "t a$ 72??<8.A 3ydrogels are formed when a three-dimensional
polymeric networ& is loosely crosslin&ed. 'hey are swollen by waterbut not dissolved in it.
3ydrogels may display reversible solGgel transitions, induced by changes in the
environmental conditions such as temperature, p3, ionic strength, phase separation, wave
length of light, crystallinity, etc. 3ydrogelis described as smart or intelligent when sharp
transition is induced by small change in such conditions. Ior the shapememoryhydrogels,
reversible change in shape may also be induced by such stimuli. 'he preparation and
applicationsof the molecularly imprinted polymeric hydrogels 7,4s" are illustrated by a few
e*amples. 'he use of shape sensitivehydrogels in microfluidic is mentioned. #pplication of
hydrogels for chronobiology and chronotherapy is outlined. 'he conversion of hydrogels into
aerogels and their respective properties is discussed.
Ma S@ "t a$ 72??;8.A 4repared a novel ocular cationic microemulsion-in situ gel C7-,;G"
system with vitamin. H#41C7 was prepared by a process based on supply of energy, and the
before-and-after gelation rheology of H#41C7-,;G was investigated. ,n vitro H#4 release
and gel dissolution of both H#41C7-,;G and (culotect Gel was determined. (cular irritation
test was carried out based on the $rai+e method. Rheology study illustrated that H#41C7
reduced the phase transition temperature of 4olo*amer .92 gel by !.? degrees C, and the
elastic modulus increased about !?.2 times. 'he in vitro release and gel dissolution profile of
both formulations e*hibited the characteristics of +ero order &inetics. Comparing with
(culotect Gel, desorption &inetics study of H#41C7-,;G e*hibited longer corneal retention
time and smaller contact angle. ,rritation test showed a good ocular compatibility of
H#41C7-,;G.
Mai Man+o(% "t a$ 72??;8.A $eveloped in situ gelling formulations of ciproflo*acin
hydrochloride 3Cl" aiming at prolonging corneal contact time, controlling drug release,
enhancing ocular bioavailability, and increasing patient compliance. 'he in situ forming gels
were prepared using different concentrations of polo*amer .92 4.92" and polo*amer !55
4!55". 7ucoadhesives such as hydro*ypropylmethyl cellulose 347C" or hydro*yethyl
cellulose 3EC" were added to the formulations to enhance the gel bioadhesion properties.
'he gelation temperature of the prepared formulations ranged from 25.99 to )..9)RC.
,ncreasing the concentrations of 4.92, 347C, and 3EC increased the viscosity and
mucoadhesion force of the preparations and decreased the in vitro drug release. Ciproflo*acin
3Cl in situ forming gel formulae composed of 4.9214!551347C !51!)1!.?>, wt1wt", and
4.9214!5513EC !51!)19.?>, wt1wt" showed optimum release and mucoadhesion properties
and improved ocular bioavailability as evidenced by an enhanced therapeutic response
compared with the mar&eted conventional eye drops.
@"n51i Ma "t a$ 72??;8.A Concluded ocular in situ gel prolong the precorneal resident time
and improve ocular bioavailability of the drug, 4luronic-g-polyacrylic acid" copolymers were
studied as a temperature-responsive in situ gelling vehicle for an ophthalmic drug delivery
system. 'he rheogram and in vitro drug release studies indicated that the drug release rates
decreased as acrylic acid14luronic molar ratio and copolymer solution concentration
increased. 'he release rates of drug from such copolymer gels were mainly dependent on the
gel dissolution. ,n vivo resident e*periments showed the drug resident time and the total
resident amount increased by .-fold and !.2-fold for in situ gel compared with eye drops.
Li( E "t a$ 72??:8.A $eveloped poor bioavailability and therapeutic response e*hibited by
conventional ophthalmic solutions due to rapid precorneal elimination of the drug may be
overcome by the use of gel system, the evaluation was conducted to &now the relative
bioavailability of ion-activated in situ ophthalmic gel of gatiflo*acin by microdialysis.
Mat,(% R "t a$ 72??:8.A $eveloped formulation and evaluation of an ocular delivery system
of timolol maleate based on the concept of both temperature and p3-triggered in situ gelation.
4luronic I-!22 a thermosensitive polymer" in combination with chitosan p3-sensitive
polymer also acts as permeation enhancer" was used as gelling agent.
@ang C "t a$ 72??:8.A Reported formulation shows the precorneal resident time and improve
ocular bioavailability of the drug, 4luronic I!22-g-polyacrylic acid" copolymers were
studied as in situ gelling vehicle for ophthalmic drug delivery system.'he rheogram and in
vitro drug release studies indicated that the drug release rates decreased as acrylic
acid14luronic molar ratio and copolymer solution concentration increased. 'he release rates of
the drug from such copolymer gels were mainly dependent on the gel dissolution.
Yan>ia Cao

"t a$ 72??:8.A ,nvestigated poly L-isopropylacrylamide"-chitosan 4L,4##m-
C;", was investigated for its thermosensitive in situ gel-forming properties and potential
utili+ation for ocular drug delivery. 'he in vivo ocular pharmaco&inetics of timolol maleate in
4L,4##m-C; solution were evaluated and compared to that in conventional eye drop
solution by using rabbits according to the microdialysis method Iurthermore, the 4L,4##m-
C; gel-forming solution of timolol maleate had a stronger capacity to reduce the intra-ocular
pressure ,(4" than that of the conventional eye drop of same concentration over a period of
!2 h. 'hese results suggest that 4L,4##m-C; is a potential thermosensitive in situ gel-
forming material for ocular drug delivery, and it may improve the bio-availability, efficacy,
and compliance.
&a+a)a%a' 0. Nan'a-a1" "t a$ 72??:8.A (phthalmic drug delivery is one of the most
interesting and challenging endeavors facing the pharmaceutical scientist. 'he conventional
oculardrug delivery systems li&e solutions, suspensions, and ointments show drawbac&s such
as increased precorneal elimination, high variability inefficiency, and blurred vision
respectively. ,n situ-forming hydrogels are li%uid upon instillation and undergo phase
transition in the ocular cul-desac to form visco-elastic gel and this provides a response to
environmental changes. ,n the past few years, an impressive number of noveltemperature, p3,
and ion induced in situ-forming systems have been reported for sustain ophthalmic drug
delivery. Each system has its ownadvantages and drawbac&s. 'he choice of a particular
hydrogel depends on its intrinsic properties and envisaged therapeutic use. 'his
reviewincludes various temperature, p3, and ion induced in situ-forming polymeric systems
used to achieve prolonged contact time of drugs with the cornea and increase their
bioavailability.
C,(n'i" @U "t a$ 72??98.A $eveloped p3-triggered in situ gelling vehicle for ophthalmic
delivery of puerarin, the effect of hydro*ypropyl-6-cyclode*trin 34-6-C$" on the a%ueous
solubility and in vitro corneal permeation of puerarin was also investigated. 'he puerarin
solubility increased linearly and proportionally to the 34-6-C$ concentrations and ?> w1v"
34-6-C$ enhanced its ocular permeability significantly. Carbopol 859LI was used as the
gelling agent in combination with 347C 7ethocel E.7" which acted as a viscosity-
enhancing agent. 'he optimum concentrations of Carbopol 859LI and 347C E.7 for the in
situ gel-forming delivery systems were 9.!> w1v" and 9..> w1v", respectively.
E,i1ong Li( "t a$ 72??98.A Concluded formulation and evaluation of an ophthalmic delivery
system of an antibacterial agent, gatiflo*acin, based on the concept of ion-activated in situ
gelation. #lginate Nelton" was used as the gelling agent in combination with 347C
7ethocel E?9Lv" which acted as a viscosity-enhancing agent. 'he rheological behaviors of
all formulations were not affected by the incorporation of gatiflo*acin. Both in vitro release
studies and in vivo pre-corneal retention studies indicated that the alginate1347C solution
retained the drug better than the alginate or 347C E?9Lv solutions alone.
0. C. S(ng "t a$ 72??38.A ,nvestigated alginate and 4luronic-based solutions as the in situ
gelling vehicles for ophthalmic delivery of pilocarpine, the optimum concentration of alginate
solution for the in situ gel-forming delivery systems was 2> w1w" and that for 4luronic
solution was !.> w1w". 'he mi*ture of 9.!> alginate and !.> 4luronic solutions showed a
significant increase in gel strength in the physiological condition- this gel mi*ture was also
found to be free flowing at p3 ..9 and 2? RC.
S(1i.ta Gang($ "t a$ 72??38.A 'he ob0ective of this study was to develop a novel chitosan-
glyceryl monooleate G7(" in situ gel system for sustained drug delivery and targeting. 'he
delivery system consisted of )> w1v" chitosan and )> w1v" G7( in 9.)) 7 citric acid. ,n
situ gel was formed at a biological p3. ,n vitro release studies were conducted in ;orensenAs
phosphate buffer p3 2.." and drugs were analy+ed either by 34LC or spectrophotometry.
7ucoadhesive property of the gel was evaluated in vitro using an EQ-'ester. ,ncorporation of
a cross-lin&er glutaraldehyde" retarded the rate and e*tent of drug release.
@ata%( 0(4o "t a$

72??28.A (ral administration of a%ueous solutions of either gellan gum
!.9> w1v" or sodium alginate !.?> w1v" containing calcium ions in comple*ed form
resulted in the formation of gel depots in rabbit and rat stomachs as a conse%uence of the
release of the calcium ions in the acidic environment. ,n vitro studies demonstrated diffusion-
controlled release of paracetamol from the gels over a period of /h. 'he bioavailability of
paracetamol from the gels formed in situ in the stomachs of rabbits following oral
administration of the li%uid formulations was similar to that of a commercially available
suspension containing an identical dose of paracetamol.
N. 0a-a+aDi "t a$ 72??18.A $eveloped thermoreversible gels formed in situ by a%ueous
solutions of an en+yme-degraded *yloglucan polysaccharide were evaluated as sustained
release vehicles for the ocular delivery of pilocarpine hydrochloride. 'he miotic responses in
rabbit following administration of *yloglucan sols were compared with those from in situ
gelling 4luronic I!22 sols and from an a%ueous buffer solution containing the same drug
concentration. ;ustained release of pilocarpine was observed with all gels, the duration of
miotic response increasing with increase of *yloglucan concentration.
P.D. Amin "t a$ 72??18.A 'he poor bioavailability and therapeutic response e*hibited by
conventional ophthalmic solution due to rapid pre corneal elimination of the drug may be
over come by the use of in situ gel forming system that are instilled as drops into the eye and
under go sol-gel in the cu-de-sac.the present wor& describes the formulation and evaluation of
ophthalmic in situ system of an anti bacterial agent (flo*acin based on the concept of p3
triggered in situ gelation.carbopol-8.9 was used as gelling agent in combination with 347C
E ?9 LH which acted as a viscosity enhancing agent .the developed formulation was
therapeutically efficacious , stable ,non irritant and provided sustained release of drug over an
5hr .
Y(t,iDa H. Sama%anaaD" "t a$ 72??18.A 'here are many animal models for evaluating anti
fungal activity. 'he mon&ey, rat, and mouse are

the choice models for investigating oral
candidiasis, but comparisons

between the same or different models appear difficult, because

of
variables such as the study design, the number of animals used,

their diet, the differences in
Candida strains, and the duration

of the studies. 'hese variables notwithstanding, the
following

could be concluded. i" 'he primate model is ideal for investigating

Candida-
associated denture stomatitis since both erythematous

and pseudomembranous lesions have
been produced in mon&eys with

prosthetic plates- they are, however, e*pensive and difficult

to
obtain and maintain. ii" 'he rat model both ;prague-$awley

and Cistar" is well proven for
observing chronic oral candidal

coloni+ation and infection, due to the ease of breeding and
handling

and their ready availability. iii" 7ice are similar, but in addition

there are well
characteri+ed variants simulating immunologic and

genetic abnormalities e.g., athymic,
euthymic, murine-ac%uired

immune deficiency syndrome, and severe combined
immunodeficient

models" and hence are used for short-term studies relating the

host immune
response and oral candidiasis. Lonetheless, an ideal,

relatively ine*pensive model
representative of the human oral

environment in ecological and microbiological terms is yet to
be described. Entil such a model is developed, researchers should

pay attention to
standardi+ation of the e*perimental protocols

described here to obtain broadly comparable and
meaningful

data.
Ya+(Di Damai "t a$ 72??18.A # straightforward murine model of oropharyngeal candidiasis.
7ice were immune suppressed with cortisone acetate, anestheti+ed, and then inoculated by
placing cotton wool balls saturated with Candidaalbicans sublingually for 2 h. # prolonged,
reproducible infection was induced. 'his model may be useful for antifungal screening or
pathogenesis studies.
E,ang // "t a$ 72???8.A $eveloped flucona+ole ICQ" in the a%ueous humors and tears of
Lew Qealand white rabbits following topically applied in-situ-forming gels of 9.?> ICQ
,;G-ICQ" and 9.?> ICQ eye drops to rabbit eyes. 'he rabbit tears and a%ueous humors were
obtained and %uantified at different times after topically applying single dose of ,;G-ICQ and
ICQ eye drops to the rabbit eyes. ICQ concentrations in rabbit tears within !59 min after
applying ,;G-ICQ were found to be significantly higher compared with those of ICQ eye
drops.
T,oma+ '. @a$+, "t a$ 72???8.A (ropharyngeal and esophageal candidiasis (4EC" is a
fre%uent opportunistic mycosis in immunocompromised patients. #+ole-resistant (4EC is a
refractory form of this infection occurring particularly in human immunodeficiency virus
3,H"-infected patients. 'he procedures developed by the #ntifungal ;ubcommittee of the
Lational Committee for Clinical Laboratory ;tandards LCCL;" are an important advance in
standardi+ation of in vitro antifungal susceptibility methodology. ,n order to further
understand the relationship between LCCL; methodology and antifungal therapeutic
response, we studied the potential correlation between in vitro susceptibility to flucona+ole
and in vivo response in a rabbit model of flucona+ole-resistant (4EC. 7,Cs of flucona+ole
were determined by LCCL; methods.
'hree flucona+ole-susceptible I;" 7,C, U9.!2? mg1ml" and three flucona+ole-
resistant IR" 7,C, O/. mg1ml" isolates of Candida albicans from prospectively monitored
3,H-infected children with (4EC were studied. IR isolates were recovered from children
with severe (4EC refractory to flucona+ole, and I; isolates were recovered from those with
mucosal candidiasis responsive to flucona+ole. Ilucona+ole at 2 mg1&g of body weight1day
was administered to infected animals for 2 days. 'he concentrations of flucona+ole in plasma
were maintained above the 7,Cs for I; isolates throughout the dosing interval. Ilucona+ole
concentrations in the esophagus were greater than or e%ual to those in plasma. Rabbits
infected with I; isolates and treated with flucona+ole had significant reductions in oral
mucosal %uantitative cultures 4 U 9.99!" and tissue burden of C. albicans in tongue, soft
palate, and esophagus 4 U 9.99!". ,n comparison, rabbits infected with IR isolates were
unresponsive to flucona+ole and had no reduction in oral mucosal %uantitative cultures or
tissue burden of C. albicans versus untreated controls. Ce conclude that there is a strong
correlation between in vitro flucona+ole susceptibility by LCCL; methods and in vivo
response to flucona+ole therapy of (4EC due to C. albicans.
/o,an Ca%$!o%+ "t a$ 71<<;8.A Reported Gelritein situ gels in vivo study for determining
precorneal contact times in humans and in rabbits was performed. 'he elastic moduli of the
gels increased with increasing concentration of electrolytes. #t physiological concentration of
the electrolytes, the elasticity of the gels was independent of Gelrite concentration. 'he
human contact times increased up to 29 h with decreasing osmolality of the formulations. 'he
results indicate that a high rate of the sol1gel transition results in long contact times.
0ata%ina E1+man "t a$ 71<<:8.A ;tudied rheological measurements were performed to study
the gel and the solGgel transition of an in situ gel, 4olo*amer .92. #n increasing
concentration of polo*amer resulted in a slightly increasing elasticity of the gels and a
decreasing solGgel transition temperature. 'he contact time increased with increasing
concentration of polo*amer which could be e*plained and correlated with the rheology of
polo*amer solutions1gels mi*ed with simulated tear fluid. 'he polo*amer system did not
seem to be promising as an ophthalmic in situ gel due to the strong concentration dependence
of the solGgel transition temperature combined with the dilution that occurs in the eye.
Chapter-) ;cope (f Cor&
2. SCOPE OF @OR0
Conventional opthalmic solutions result in poor bio availability and theraputic
response because high tear fluid turn over and dynamic cause rapid pre corneal elimination of
drug. # high fre%uency of eye drops installation associated with patient non complience.
'he low bioavailability of drug from an ophthalmic solution is attributed to naso
lachrymal drynage in con0unsction with low drug permeability of cornea. 'he rapid
elimination of instilled drug often results in short duration of theraputic effect and
conse%uently the need for fre%uent dosing regimen.
# significant increase in pre corneal residence time of drug and conse%uently increase
in bioavailability can be achieved by using delivery systems based on concept of in situ gel
formation. 'hese systems consist of polymers that e*hibit sol to gel phase transition due to
change in specific physico chemical parameters in their environment. 'he sol-gel transition
can be induced by a shift in p3, temperature or ion activated systems.
Gatiflo*acin, a fourth generation /-fluro-5-metho*y %uinolone has a broader anti
bacterial spectrum against gram positive and gram negative organisms. 'hese
fluoro%uinolones are routinely used in management of of ocular infections li&e severe
bacterial &eratitis, endopthalmitis, con0unctivitis, corneal ulcers etc. Conventional ophthalmic
dosage form of gatiflo*acin has inherent draw bac&s li&e naso lacrimal drainage, fre%uent
dosing regimen, less contact time, poor bio availability and patient non-compliance.
Chloramphenicol has a broader anti bacterial spectrum against gram positive and
gram negative organisms. 'hese fluoro%uinolones are routinely used in management of of
ocular infections li&e severe bacterial &eratitis, endopthalmitis, con0unctivitis, corneal ulcers
etc. Conventional ophthalmic dosage form of Chloramphenicol has inherent draw bac&s li&e
naso lacrimal drainage, fre%uent dosing regimen, less contact time, poor bio availability and
patient non-compliance.
,n the precent study, an attempt is being made to develop thermoreversible insitu
gelling ophthalmic delivery system of gatiflo*acin, chloramphenicol inorder to enhance its
therapeutic efficacy and to over come the inherent draw bac&s associated with conventional
gatiflo*acin, chloramphenicol ophthalmic formulation.
'he in situ forming gels of Gatiflo*acin, Chloramphenicol will be prepared using
thermoreverse polymer, pola*amer !55 and viscosity enhancing agent such as chitosan. 'he
prepared in situ gelling system will be evaluated for different parameters such as p3,
appearance, drug content, rheology, gelation studies, in vitro release studies, sterility, in vitro
efficacy.
Chapter-) ;cope (f Cor&
Chapter-. $rug #nd E*cipient 4rofile
3. DRUG AND EBCIPIENT PROFILE
3.1. GATIFLOBACIN
C,"mi#a$ nam":
!-cyclopropyl-/-fluoro-!,.-dihydro-5-metho*y-2)-methyle-!pypara+inyl"-.-o*o-)-
%uinoline carbo*ylic acid.
St%(#t(%a$ !o%m($a:
Fig. No. < +t%(#t(%a$ !o%m($a o! gati!$o>a#in
Em.i%i#a$ !o%m($a: C
!8
3
22
IL
)
(
.
.!?32(
Mo$"#($a% -"ig,t: .92..2 gms1mol.
D"+#%i.tion:
Gatiflo*acin is a ses%uihydrate chrystalline powder and is white to pale yellow in
colour. ,t e*ists as a racemate, with no net optical rotation. 'he solubility of compound is p3
dependent. 'he ma*imum a%ueous solubility .9-/9 mg1ml" occurs at a p3 range of 2 to ?.
2)"
A4+o%.tion:
Gatiflo*acin is well absorbed of the g.i tract after oral administration and can be
given with out regarding to food. 'he absolute bioavailability of gatiflo*acin is 8/>. 4ea&
plasma concentration of gatiflo*acin usually occurs !-2 hours after oral dosing. 2."
P,a%ma#oDin"ti#+:
Gatiflo*acin pharmaco&inetics are linear and time independent at doses ranging from
299 to 599 mg administered over a period of up to !. days. ;teady state concentrations are
achieved by third daily oral or intravenous dose of Gatiflo*acin. 'he mean steady state pea&
and trough plasma concentrations attained following a dosing regimen of .99 mg once daily
are appro*imately ..2 Dg1ml and 9.. Dg1ml respectively for oral administration and ../ Dg1ml
and 9.. Dg1ml respectively for intravenous administration.2?"
Di+t%i4(tion:
;erum protein binding of Gatiflo*acin is appro*imately 29> and is concentration
independent. 'he mean volume of distribution of Gatiflo*acin at steady state ranged from !.?
to 2 L1&g. Gatiflo*acin is widely distributed throughout the body into many body tissues and
fluids. Rapid distribution of Gatiflo*acin into tissues results in higher Gatiflo*acin
concentration in most target tissues than in serum. 2/"
M"ta4o$i+m:
Gatiflo*acin undergoes limited biotransformation in humans with less than !> of the
dose e*creted in urine as metabolites. 22"
E>#%"tion:
Gatiflo*acin is e*creted as unchanged drug primarily by the &idney. 'he mean
elimination half life of Gatiflo*acin ranges from 2 to !. hours and is independent of dose and
route of administration. 25"
O.,t,a$mi# +."#i!i#ation+:
Ior the treatement of bacterial specifications caused by susceptible microorganisms,
blepharitis blepharo con0ectivitis, &erato con0unctivitis, scleritis, episcleritis,
merbomianitis.#lso reported useful in pre and post ocular surgery li&e L#;,N, cataract
pVrocedure and other ocular surgery.
Antimi#%o4ia$ "!!i#a#:
Gatiflo*acin has shown to be active against most strans of the following micro
organisms both invitro and clinically in con0uctival infections. 28"
A"%o4"+A g%am .o+iti)":
Corneyebacterium propingum
;taphylococus aureus
;taphylo cocus epidermidis
;trepto cocus mitis
;treptococus pneumonia
#erobes, gramnegative:
3aemophilus influen+a
D%(g int"%a#tion+:
;pecific drug interaction studies have been conducted with gatiflo*acin opthalmic
solution. 3owever systemic administration of some %uinolones have been shown to elevate
pharms concentration of theophyllin, interfer with metabolism of caffeine, enhance the effect
of oral anti coagulant warfarin and its derivative s and has been associated with trancient
elevations in patient receiving systemic cyclosporine concomitantly.)9"
Cont%ain1i#ation+:
3ypersensitivity to Gatiflo*acin or other %uinolones in the medication.
A1)"%+" "!!"#t+:
7ild con0unctival irritation, increased lacrimation, occasionally eye discharge,
irritation, lidedema and headache.
Do+ag" an1 a1mini+t%ation:
'he recommended dosage regimen for the treatment of bacterial con0unctivitis is:
$ays ! and 2: ,nstill one drop every two hours in the affected eyes" while awa&e,
ma*imum up to 5 times daily.
$ays ) to 2: ,nstill one drop up to four times daily while awa&e. ,n other conditions:
#s advised by the doctor. )!"
Sto%ag":
;tore in a cool dry place, protected from light.
Ma%D"t"1 !o%m($ation+ o! Gati!$o>a#in:
Ta4$" no: 2 Ma%D"t"1 !o%m($ation+ o! Gati!$o>a#in
S%. No. &%an1 Nam" St%"ngt, Man(!a#t(%"%
! G#'E $4; 9.)> #0anta 4harma Ltd.
2 G#',N,L$ $4; 9.)>
7an&ind 4harma 4vt.
Ltd.
) G#',WE,L $4; 9.)> Cipla Ltd.
. G#'' $4; 9.)> Le ;ante
? QELG#' $4; 9.)> Nlarschen 4vt.Ltd.
/ Q,G#' $4; 9.)> I$C Ltd.
3.2. CHLORAMPHENICOL PALMITATE
C,"mi#a$ nam":
$--"-threo-!-p-nitrophenyl"-2-dichloroacetamindo-!, )-propanediol
2,2-dichloro-L-2-hydro*y-!-hydro**ymethyle"-2-.nitrophenyl"ethyl"acetamide
$-threo-L-dichloroacetyl-!-paranitrophenyl!-2-amino-!, )-propane-diol
$ -" threo-2-dichloroacetamido-!-p-nitrophenyl-propanediol.
St%(#t(%a$ !o%m($a:
Fig(%" 1?: St%(#t(%a$ !o%m($a o! #,$o%am.,"ni#o$
Em.,i%i#a$ !o%m($a: C
!!
3
!2
C
l2
L
2
(
?
Mo$"#($a% -"ig,t: )2).!)22grm1mol
D"+#%i.tion:
Chloramphenicol is a large spectrum antibiotic with antimicrobial activity. ,ts
mechanism of action is based on the inhibition of protein synthesis. 3owever, the resistance
of gram-positive and gram-negative microorganisms in vivo is a clinical problem of
increasing importance .Chloramphenicol is available for oral administration as
chloramphenicol palmitate, a prodrug of chloramphenicol, developed with the ob0ective of a
more pleasent flavored derivative. )2"
Ta4$" no: 3 P,+i#a$ an1 #,"mi#a$ .%o."%ti"+ o! #,$o%am.,"ni#o$
P%o."%t In!o%mation
P,+i#a$ +tat" an1 a.."a%an#" ;olid
O1o% (dourless
Mo$"#($a% -"ig,t )2).!) g1mole
Co$o% Chite to grayish-white or yellowish-white
Ta+t" Bitter burning
A4+o%.tion:
'he absorption of drugs from solid pharmaceutical forms after oral administration
depends, among other factors, on the liberation of the drug from the pharmaceutical form, its
dissolution or solubility in physiological conditions, and its permeability through the
gastrointestinal tract.
P,a%ma#oDin"ti#+:
Gatiflo*acin pharmaco&inetics are linear and time independent at doses ranging from
299 to 599 mg administered over a period of up to !. days. ;teady state concentrations are
achieved by third daily oral or intravenous dose of Gatiflo*acin. 'he mean steady state pea&
and trough plasma concentrations attained following a dosing regimen of .99 mg once daily
are appro*imately ..2 Dg1ml and 9.. Dg1ml respectively for oral administration and ../ Dg1ml
and 9.. Dg1ml respectively for intravenous administration.))"
Di+t%i4(tion:
;erum protein binding of Gatiflo*acin is appro*imately 29> and is concentration
independent. 'he mean volume of distribution of Gatiflo*acin at steady state ranged from !.?
to 2 L1&g. Gatiflo*acin is widely distributed throughout the body into many body tissues and
fluids. Rapid distribution of Gatiflo*acin into tissues results in higher Gatiflo*acin
concentration in most target tissues than in serum. )."
M"ta4o$i+m:
C7C is metaboli+ed into at least / metabolites: nitroso-C7C L(-C7C" which is
formed in liver. ) e*cretion products which are the glucuronide C7C-G", the C7C base
L#4$", and an alcoholic derivative, 3#4. $ehydro-C7C $3-C7C" and the dehydro-
C7C base L4#4" are formed by enterobacteria in the large bowel. Concerning about
to*icity, some of the C7C metabolites are more to*ic than the parent compound and may be
to*ic to the bone marrow. 28"
'he primary metabolite of C7C is the glucoronide con0ugate. C7C arylamide is
formed by intestinal reduction of the nitro group of C7C to amine, which is acytelated and
e*creted in the urine. 3uman liver microsomes can reduce the nitro group of C7C. (*amic
acid, o*amylethanolamine and aldehyde derivatives also have been identified as metabolites
of chloramphenicol.
O.,t,a$mi# in1i#ation+:
#pply . to / times daily to the affected eye for the first 22 hours depending upon the
severity of the condition. Continue treatment for .5 hours after the eye appears normal.
In1i#ation+:
'reatment of bacterial con0unctivitis and ocular inflammations, Caused by organisms
susceptible to chloramphenicol. )?"
Limitation+:
#s with other antibiotics, prolonged use may result in overgrowth of non-susceptible
organisms. ,f superinfection occurs or if clinical improvement is not noted within a
reasonable period, discontinue use and institute appropriate therapy. #ll topical ophthalmic
preparations containing corticosteroids, with or without an antimicrobial agent, are
contraindicated in the initial treatment of corneal ulcers. 'hey should not be used until the
infection is under control and corneal regeneration is well under way. 'his chloramphenicol
product must not be used in meat, egg or mil& producing animals. 'he length of time that
residues persist in mil& or tissues has not been determined.
P,a%ma#o$ogi# P%o."%ti"+
'he e*ertion of its bacteriostatic effect is shown by binding to the bacterial ?9;
ribosomal subunit and inhibits protein synthesis at the peptidyl transferase reaction.,t may be
administered either intravenously or orally $rug ,nformation 3andboo&, 2998". ,t is rapidly
absorbed from the gastrointestinal tract in humans and animals, with pea& values in plasma
being reached within about two to three hours of administration. ,t is completely absorbed via
the oral route and hence widely distributed throughout the body due to its lipophilic nature. ,t
readily enters the normal C;I. 'he drug inhibits the hepatic mi*ed-function o*idases.
E*cretion of the drug depends on its version in the liver to the glucuronides, which is then
secreted by the renal tubules and also secreted into breast mil& as well. ,n humans, about !?>
is eliminated as the parent compound and the remaining 2?> in the form of metabolites. )/"
A1)"%+" D%(g R"a#tion
'he ,nternational #gency for Research on Cancer ,#RC" reviewed C7C again in
!852 and in !889. 'hat agency concluded that there was limited evidence of carcinogenicity
in humans and inade%uate evidence of carcinogenicity in e*perimental animals. ,n ma&ing the
overall evaluation, the ,#RC noted that C7C induces aplastic anemia and that this condition
is related to the occurrence of leu&emia. 'he ,#RCAs overall evaluation is that C7C is
probably carcinogenic to humans. )2"
Do+" %"$at"1 a1)"%+" "!!"#t+
3aematologic
Bone marrow suppression
3aemolytic anaemia
Cardiac
Cardiovascular collapse grey baby syndrome"
Leurologic
(ptic neuritis
4eripheral neuritis
Encephalopathy
3eadache
7ental confusion, depression
(ther
3ypersensitivity reactions
Lausea, vomiting, diarrhoea
4seudomembranous colitis
Glossitis, stomatitis
(toto*icity topical otic formulations"
Not 1o+" %"$at"1
3aematologic
,diosyncratic aplastic anaemia
'hese adverse effects have led to dramatic reductions in the use of chloramphenicol
in developed country settings. Ese of chloramphenicol has been replaced by other antibiotics
with a better side effect profile, although it remains a useful agent. ,n resource poor settings,
its use has continued because of its availability and low cost. )5"
Ta4$" no: 6 A1)"%+" %"a#tion+ o! #,$o%am.,"ni#o$
Cat"go% A1)"%+" %"a#tion+
C"nt%a$ n"%)o(+ ++t"m Confusion, delirium, depression, fever, headache
D"%mato$ogi# #ngioedema, rash, urticaria
Ga+t%oint"+tina$ $iarrhea, enterocolitis, glossitis, nausea, stomatitis, vomiting
H"mato$ogi#
#plastic anemia, bone marrow suppression,
Granulocytopenia, hypoplastic anemia, pancytopenia,
thrombocytopenia
O#($a% (ptic neuritis
Mi+#"$$an"o(+ #naphyla*is, hypersensitivity reactions, Gray syndrome
Sto%ag":
;tore in a cool dry place, protected from light.
Po$m"%+ (+"1 !o% in +it( g"$ation:
Lumbers of polymers are capable of prolonging the residence time of drug delivery
systems by their in-situ gelling properties. Ior e*ample, chitosan is used in eye drops
'imoptic XE- CibaHision". ;uch in-situ gelling properties are also beneficial for numerous
other drug delivery systems improving the bioavailability and prolonging the drug action.

,n
comparison to so far used in-situ gelling polymers, thiolated polymers are capable of
providing a comparatively more pronounced increase in viscosity after application, as an
e*tensive cross lin&ing process by the formation of disulfide bonds between the polymer
chains ta&es place.

Iormation of inter- and intramolecular disulfide bonds within the
polymeric networ& stabili+es drug delivery systems based on thiomers. (ther polymers used
in the formulations are polyacrylic acid polymers li&e carbopol, chitosan and its derivatives,
polyo*ypropulene-polyo*yethylene bloc& copolymers, polo*amer !55.
3.2. CAR&APOL
Ca%4o.o$ .o$m"%+ are polymers of acrylic acid cross-lin&ed with polyal&enyl ethers
or divinyl glycol. 'hey are produced from primary polymer particles of about 9.2 to /.9
micron average diameter. 'he flocculated agglomerates cannot be bro&en into the ultimate
particles when produced. Each particle can be viewed as a networ& structure of polymer
chains interconnected via cross-lin&ing.
Ca%4om"%+ were first prepared and patented in !8?2. ;ince then, a number of
e*tended release tablet formulations, which involve carbomer matrices, have been patented
)8".
Carbomers readily absorb water, get hydrated and swell. ,n addition to its hydrophilic
nature, its cross-lin&ed structure and its essentially insolubility in water ma&es Carbopol a
potential candidate for use in controlled release drug delivery system .9".
C,"mi#a$ nat(%":
'he E;4-LI, European 4harmacopoeia, British 4harmacopoeia, Enited ;tates
#dopted Lames Council E;#L", and ,nternational Lomenclature for Cosmetic ,ngredients
,LC," have adopted the generic i.e., non-proprietary" name JcarbomerK for various
Carbopol homopolymer polymers. 'he Yapanese 4harmacopoeiastates, Carbopol
homopolymers as Jcarbo*yvinyl polymerK and Jcarbo*y polymethylene.K 'he ,talian
4harmacopoeia also identifies Carbopol 8).4 as Jcarbo*y polymethyleneK and the $eutschen
#rt+neibuch calls Carbopol 859LI Jpolyacrylicacid.K Carbopol copolymers, such as
Carbopol !).2 LI and !)52, have also been named JcarbomerK by the E;4-LI, but are
considered J#crylates1C!9-C)9 #l&yl #crylates Cross polymerK by the ,LC,.
Ca%4o.o$ .o$m"%+ are offered as fluffy, white, dry powders !99> effective".
'he carbo*yl groups provided by the acrylic acid bac&bone of the polymer are
responsible for many of the product benefits. Carbopol polymers have an average e%uivalent
weight of 2/ per carbo*yl group. 'he general structure
can be illustrated with fig. Lo.!!
Fig. No. 11 G"n"%a$ St%(#t(%"+ o! Ca%4o.o$ Po$m"%+.
Fig. No. 12 S#,"mati# 1%a-ing o! a mo$"#($a% +"gm"nt o! a #%o++5
$inD"1 .o$m"%
Po$a#%$i# a#i1 .o$m"%:
Carbopol polymers are manufactured by cross-lin&ing process. $epending upon the
degree of cross-lin&ing and manufacturing conditions, various grades of Carbopol are
available. Each grade is having its significance for its usefulness in pharmaceutical dosage
forms.
Carbopol 8). 4 is cross-lin&ed with allyl sucrose and is polymeri+ed in solvent
ben+ene.
Carbopol 2!G, 82! 4, 82. 4 are cross-lin&ed with allyl penta erythritol and
polymeri+ed in ethyl acetate. 4olycarbophil is cross-lin&ed polymer in divinyl glycol and
polymeri+ed in solvent ben+ene. #ll the polymers fabricated in ethyl acetate are neutrali+ed
by !-)> potassium hydro*ide. 'hough Carbopol 82! 4 and Carbopol 82. 4 are manufactured
by same process under similar conditions, the difference in them is that Carbopol 82! 4 has
slightly lower level of cross-lin&ing agent than Carbopol 82. 4. Carbopol 2! G is the granular
form Carbopol grade .!".
P,+i#a$ P%o."%ti"+ 7328:
'he three dimensional nature of these polymers confers some uni%ue characteristics,
such as biological inertness, not found in similar linear polymers. 'he Carbopol resins are
hydrophilic substances that are not soluble in water. Rather, these polymers swell when
dispersed in water forming a colloidal, mucilage-li&e dispersion. .)"
Carbopol polymers are bearing very good water sorption property. 'hey swell in
water up to !999 times their original volume and !9 times their original diameter to form a
gel when e*posed to a p3 environment above ..9 to /.9. Because the pNa of these polymers
is /.9 to 9.?, the carbo*ylatemoiety on the polymer bac&bone ioni+e, resulting in repulsion
between the native charges, which adds to the swelling of the polymer. 'he glass transition
temperature of Carbopol polymers is !9?RC 22!RI" in powder form. 3owever, glass
transition temperature decreases significantly as the polymer comes into contact of water. 'he
polymer chains start gyrating and radius of gyration becomes increasingly larger.
7acroscopically, this phenomenon manifests itself as swelling.
Ta4$" No.9: P,+i#a$ an1 C,"mi#a$ P%o."%ti"+ o! Ca%4o.o$ 7<238
#ppearance Iluffy, white, mildly acidic polymer
Bul& $ensity #ppro*imately 295 &g1m
)
!) lbs. ft
)
"
;pecific gravity !..!
7oisture content 2.9> ma*imum
E%uilibrium moisture content 5-!9> at ?9> relative humidity"
4Na /.9 F 9.?
p3 of !.9> water dispersion 2.? - ).9
p3 of 9.?> water dispersion 2.2 - ).?
E%uivalent weight 2/ F .
#sh content 9.998 ppm average" ZZ
Glass transition temperature !99-!9?C 2!2-22!I"
Z 4olymers produced in co solvent a cyclohe*ane 1 ethyl acetate mi*ture" have a
bul& density of !2/ &g1m
)
!! lbs1ft
)
".
Z Z 4olymers produced in ethyl acetate have ash content as potassium sulfate" of !-
)> on average.
#pplications of Carbopol polymers:
'he readily water-swellable Carbopol polymers are used in a diverse range of
pharmaceutical applications to provide:
Controlled release in tablets.
Bioadhesion in buccal ..", ophthalmic, intestinal, nasal, vaginal .?" and
rectal applications.
'hic&ening at very low concentrations to produce a wide range of viscosities and
flow properties in topical, lotions, creams and gels, oral suspensions and transdermal gel
reservoirs.
3.3. POLOBAMER
Non.%o.%i"ta% Nam"+:
B4: 4olo*amers
4hEur: 4olo*amera
E;4LI: 4olo*amer
Snonm+:
Lutrol, 7onolan, 4luronic, polo*al&ol, polyethyleneGpropylene glycol copolymer,
polyo*yethyleneGpolyo*ypropylene copolymer, ;upronic, ;ynperonic.
C,"mi#a$ Nam" an1 CAS R"gi+t% N(m4"%:
a-3ydro-o-hydro*ypoly o*yethylene" poly o*ypropylene" poly o*yethylene"
bloc&s copolymer V899)-!!-/[.
Em.i%i#a$ Fo%m($a an1 Mo$"#($a% @"ig,t:
'he polo*amer polyols are a series of closely related bloc& copolymers of ethylene
o*ide and propylene o*ide conforming to the general formula 3( C
2
3
.
(" a C
)
3
/
(" b
C
2
3
.
(" a3.
St%(#t(%a$ Fo%m($a:
Fig. No. 12 G"n"%a$ +t%(#t(%" o! .o$a>oma%
A..$i#ation+ in P,a%ma#"(ti#a$ Fo%m($ation o% T"#,no$og:
4olo*amers are nonionic polyo*yethyleneGpolyo*ypropylene copolymers used
primarily in pharmaceutical formulations as emulsifying or solubili+ing agents. .?" 'he
polyo*yethylene segment is hydrophilic while the polyo*ypropylene segment is hydrophobic.
#ll of the polo*amers are chemically similar in composition, differing only in the relative
amounts of propylene and ethylene o*ides added during manufacture.
'heir physical and surface-active properties vary over a wide range and a number of
different types are commercially available. 4olo*amers are used as emulsifying agents in
intravenous fat emulsions, and as solubili+ing and stabili+ing agents to maintain the clarity of
eli*irs and syrups. 4olo*amers may also be used as wetting agents in ointments, suppository
bases, and gels and as tablet binders and coatings. 4olo*amer !55 has also been used as an
emulsifying agent for fluorocarbons used as artificial blood substitutes and in the preparation
of solid-dispersion systems more recently, polo*amers have found use in drug-delivery
systems. ./"
'herapeutically, polo*amer !55 is administered orally as a wetting agent and stool
lubricant in the treatment of constipation. 4ola*omer is usually used in combination with a
la*ative such as danthron. 4olo*amers may also be used therapeutically as wetting agents in
eye-drop formulations, in the treatment of &idney stones, and as s&in-wound cleansers.
D"+#%i.tion:
4olo*amers generally occur as white, wa*y, free-flowing prilled granules, or as cast
solids. 'hey are practically odorless and tasteless. #t room temperature, polo*amer !2.
occurs as a colorless li%uid.
T.i#a$ P%o."%ti"+
#cidity1al&alinity: p3 \ ?.9G2.. for a 2.?> w1v a%ueous solution.
Cloud point: O!995C for a !> w1v a%ueous solution, and a !9> w1v a%ueous solution of
polo*amer !55.
D"n+it: !.9/ g1cm ) at 2?5
9
C
F$a+, .oint: 2/95
9
C
F$o-a4i$it: solid polo*amers are free flowing.
HL& )a$(": 9.?G)9- 28 for polo*amer !55.
M"$ting .oint:
!/5
9
C for polo*amer !2.
?2G?25
9
C for polo*amer !55
.85
9
C for polo*amer 2)2
?25
9
C for polo*amer ))5
?2G?25
9
C for polo*amer .92.
Moi+t(%" #ont"nt:
4olo*amers generally contain less than 9.?> w1w water and are hygroscopic only at
relative humidity greater than 59>.
S(%!a#" t"n+ion:
!8.5mL1m !8.5 dynes1cm" for a 9.!> w1v a%ueous polo*amer !55 solution at 2?5
9
C
2..9mL1m 2..9 dynes1cm" for a 9.9!> w1v a%ueous polo*amer !55 solution at 2?5C-
2/.9mL1m 2/.9 dynes1cm" for a%ueous polo*amer solution at 2?5
9
C.
Vi+#o+it 71nami#": !999 m4a s !999 c4" as a melt at 225C for polo*amer !55.

Sta4i$it an1 Sto%ag" Con1ition+:
4olo*amers are stable materials. #%ueous solutions are stable in the presence of
acids, al&alis, and metal ions. 3owever, a%ueous solutions support mold growth.
'he bul& material should be stored in a well-closed container in a cool, dry place.
In#om.ati4i$iti"+:
$epending on the relative concentrations, polo*amer !55 is incompatible with
phenols and parabens.
M"t,o1 o! Man(!a#t(%":
4olo*amer polymers are prepared by reacting propylene o*ide with propylene glycol
to form polyo*ypropylene glycol. Ethylene o*ide is then added to form the bloc& copolymer.
Sa!"t:
4olo*amers are used in a variety of oral, parenteral, and topical pharmaceutical
formulations and are generally regarded as nonto*ic and nonirritant materials. 4olo*amers are
not metaboli+ed in the body.
#nimal to*icity studies, with dogs and rabbits, have shown polo*amers to be
nonirritating and nonsensiti+ing when applied in ?> w1v and !9> w1v concentration to the
eyes, gums, and s&in.
,n a !.-day study of intravenous administration at concentrations up to 9.? g1&g1day
to rabbits, no overt adverse effects were noted. # similar study with dogs also showed no
adverse effects at dosage levels up to 9.? g1&g1day. ,n a longerterm study, rats fed )> w1w or
?> w1w of polo*amer in food for up to 2 years did not e*hibit any significant symptoms of
to*icity. 3owever, rats receiving 2.?> w1w of polo*amer in their diet showed some decrease
in growth rate.Lo hemolysis of human blood cells was observed over !5 hours at 2?5
9
C, with
9.99!G!9> w1v polo*amer solutions. .2"
3.6. CHITOSAN
Snonm+:
beta-!,.-4oly-$-glucosamine
C992).
Chicol
Chitopearl )?!9
Chitopearl BC )999
C,"mi#a$ Nam" an1 CAS R"gi+t% N(m4"%:
4oly beta-!,.-$-glucosamine". 89!2-2/-.
St%(#t(%a$ Fo%m($a:
Fig(%": 13 G"n"%a$ +t%(#t(%" o! #,ito+an
F(n#tiona$ Cat"go%:
Chitosan is used as $ispersing agent, emulsifying, coemulsifying agent, solubili+ing
agent, tablet lubricant and wetting agent.
P,+i#a$ an1 C,"mi#a$ P%o."%ti"+:
Ta4$" No.:: P,+i#a$ an1 C,"mi#a$ P%o."%ti"+ o! #,ito+an
C,ito+an S."#i!i#ation
#ppearance Chite
4article 59 mesh powder
7oisture ] !9.9>
#sh ] !.9>
,nsoluble ] !.9>
$eacetylation $#C" ^ 5?>,89>,8?>
Hiscosity ]!99!>C';,!> 3#C,2?
9
C"
p3 value 2.9-8.9
4bppm" ] !9.9
#s ppm" ]9.?
'otal plate count ]!9991g
E.coli #bsent
Germs Lo finding
4ac&ing !912? &g
D"+#%i.tion:
Chitosan is a biopolymer derivative of chitin, a polysaccharide found abundantly in
nature in crustacean e*os&eletons of crab, shrimp, and lobster as well as in cuttlebone of
cuttlefish. ,t is a linear chain of lin&ed 2-acetoamido-2 deo*y-6-$-glycopyranose units.
Chitosan possesses uni%ue functional and biomedical properties. ,t is biodegradable,
biocompatible, nonto*ic, heat resistant showing adsorption properties and is easily accessible
by recovering from marine waste .5". ,t is especially attractive due to its film forming
characteristic and finds multiple uses in applications of coatings, drug delivery, nutrients,
controlled release of food ingredients, separation techni%ues, optical material, and so forth.
Man(!a#t(%" an1 .%o."%ti"+
Chitosan is produced commercially by deacetylation of chitin , which is the structural
element in the e*os&eleton of crustaceans crabs, shrimp, etc." and cell walls of fungi. 'he
degree of deacetylation >$$" can be determined by L7R spectroscopy, and the >$$ in
commercial chitosans is in the range /9-!99 >. (n average, the molecular weight of
commercially produced chitosan is between )599 to 29,999 daltons. # common method for
the synthesis of chitosan is the deacetylation of chitin using sodium hydro*ide in e*cess as a
reagent and water as a solvent. 'his reaction pathway, when allowed to go to completion
complete deacetylation" yields up to 85> product. .8"
'he amino group in chitosan has a pNa value of _/.?, which leads to a protonation
in acidic to neutral solution with a charge density dependent on p3 and the >$#-value. 'his
ma&es chitosan water soluble and a bioadhesive which readily binds to negatively charged
surfaces such as mucosal membranes. Chitosan enhances the transport of polar drugs across
epithelial surfaces, and is biocompatible and biodegradable. 4urified %ualities of chitosans are
available for biomedical applications.
Chitosan and its derivatives such as trimethylchitosan where the amino group has
been trimethylated" have been used in non-viral gene delivery. 'rimethylchitosan, or
%uaternised chitosan, has been shown to transfect breast cancer cells- with increased degree of
trimethylation increasing the cytoto*icity and at appro*imately ?9> trimethylation the
derivative is the most efficient at gene delivery. (ligomeric derivatives )-/ &$a" are
relatively non-to*ic and have good gene delivery properties. ?9"
A..$i#ation+ in P,a%ma#"(ti#a$ Fo%m($ation o% T"#,no$og:
a8 Nat(%a$ 4io#ont%o$ an1 "$i#ito%
,n agriculture, chitosan is used primarily as a natural seed treatment and plant growth
enhancer, and as an ecologically friendly biopesticide substance that boosts the innate ability
of plants to defend themselves against fungal infections. ?!"
Chitosan applications for plants and crops are regulated by the E4#, and the E;$#
Lational (rganic 4rogram regulates its use on organic certified farms and crops. E4#
approved biodegradable chitosan products are allowed for use outdoors and indoors on plants
and crops grown commercially and by consumers. ?2"
'he natural biocontrol ability of chitosan should not be confused with the effects of
fertili+ers or pesticides upon plants or the environment. Chitosan active biopesticides
represent a new tier of cost effective biological control of crops for agriculture and
horticulture. ?)"
'he biocontrol mode of action of chitosan elicits natural innate defense responses
within plant to resist insects, pathogens, and soil-borne diseases when applied to foliage or the
soilChitosan increases photosynthesis, promotes and enhances plant growth, stimulates
nutrient upta&e, increases germination and sprouting, and boosts plant vigor. Chen used as
seed treatment or seed coating on cotton, corn, seed potatoes, soybeans, sugar beets, tomatoes,
wheat and many other seeds, it elicits an innate immunity response in developing roots which
destroys parasitic cyst nematodes without harming beneficial nematodes and organisms.?."
#gricultural applications of chitosan can reduce environmental stress due to drought and soil
deficiencies, strengthen seed vitality, improve stand %uality, increase yields, and reduce fruit
decay of vegetables, fruits and citrus crops. ??"
48 Ho%ti#($t(%a$ a..$i#ation+:
3orticultural applications of chitosan are blooms and e*tend the life of cut flowers
and Christmas trees. 'he E; Iorest ;ervice has conducted research on chitosan to control
pathogens in pine trees and chitosan=s ability to increase pine tree resin pitch outflow by .9>
to resist pine beetle infestation. ?/"

#8 @at"% Fi$t%ation
Chitosan can also be used in water processing engineering as a part of a filtration
process. Chitosan causes the fine sediment particles to bind together, and is subse%uently
removed with the sediment during sand filtration. Chitosan also removes phosphorus, heavy
minerals, and oils from the water. Chitosan is an important additive in the filtration process.
;and filtration apparently can remove up to ?9> of the turbidity alone, while the chitosan
with sand filtration removes up to 88> turbidity. ?2" Chitosan has been used to precipitate
caseins from bovine mil& and cheese ma&ing. ?5"
Chitosan is also useful in other filtration situations, where one may need to remove
suspended particles from a li%uid. Chitosan, in combination with bentonite, gelatin, silica gel,
isinglass, or other fining agents is used to clarify wine, mead, and beer. #dded late in the
brewing process, chitosan improves flocculation, and removes yeast cells, fruit particles, and
other detritus that cause ha+y wine. Chitosan combined with colloidal silica is becoming a
popular fining agent for white wines, because chitosan does not re%uire acidic tannins found
primarily in red wines" with which to flocculate. ?8"
18 Pot"ntia$ in1(+t%ia$ (+"
;cientists have recently developed a polyurethane coating that heals its own scratches
when e*posed to sunlight, offering the promise of scratch-free cars and other products. 'he
self-healing coating uses chitosan incorporated into traditional polymer materials, such as
those used in coatings on cars to protect paint. Chen a scratch damages the chemical
structure, the chitosan responds to ultraviolet light by forming chemical chains that begin
bonding with other materials in the substance, eventually smoothing the scratch. 'he process
can ta&e less than an hour. /9"
7are& C. Erban, a scientist wor&ing on this pro0ect, said the polymer can only repair
itself in the same spot once, and would not wor& after repeated scratches?/"Chether this
technology can be applied to industrial materials, however, depends on a number of factors
long-term persistence of <healability<, stiffness and heat resistance of coating, &nowledge of
the e*act mechanism of healing, etc." not present initial studies- further investigation into
these factors can potentially ta&e decades to rectify./!"
"8 &iom"1i#a$ (+"+
Chitosan is used to rapidly clot blood, and has recently gained approval in the Enited
;tates and Europe for use in bandages and other hemostatic agents. Chitosan hemostatic
products have been shown in testing by the E.;. 7arine Corps to %uic&ly stop bleeding, and
result in !99> survival of otherwise lethal arterial wounds in swine and to reduce blood loss.
Chitosan hemostatic products reduce blood loss in comparison to gau+e dressings and
increase patient survival. /2" Chitosan hemostatic products have been sold to the E.;. #rmy
and are currently used by the EN military. Both the E; and EN have already used the
bandages on the battlefields of ,ra% and #fghanistan. Chitosan is hypoallergenic and has
natural antibacterial properties, which further support its use in field bandages. /)"
!8 C$aim"1 ,"a$t, 4"n"!it+
Chitosan is fre%uently sold in tablet form at health stores as a <fat binder<: ,t is
supposed to have the capability to interact with lipids fat" from the digestive system and limit
their absorption in the body. 'herefore, chitosan can be an effective complement to help lose
weight during diet period or to stabilise one=s weight. ,n the 2992 Cochrane meta-analysis
/." which evaluated all available clinical trials performed with chitosan on the sub0ect of
weight loss, it was concluded that body weight and all parameters related to cholesterol
changed in favor of chitosan compared to placebo. 'he various %ualities in terms of duration,
sample si+e, doses, sub0ect characteristics, type of diet, chitosan %uality and characteristics,
etc." of the clinical trials performed to evaluate the effect of chitosan on body weight might
account for some of the disparities observed in clinical trial results and the subse%uent critics
regarding the real efficacy of chitosan. ,n an e*perimental model of the stomach and
duodenum tract, chitosan has shown to interact with oil, which inhibited duodenal absorption
and enhanced lipid e*cretion. /?"
g8 M"1i#a$ R"+"a%#,
Chitosan is currently the focus of much medical research, as it is a polyglucosamine the
second-most-common dietary fiber, after cellulose".//" ;tudies have shown chitosan has the
following properties:
#s a soluble dietary fiber, it increases gastrointestinal lumen viscosity and slows
down the emptying of the stomach.
,t alters bile acid composition, increasing the e*cretion of sterols and reducing the
digestibility of ileal fats. ,t is unclear how chitosan does this, but the currently
favored hypotheses involve the increase of intestinal viscosity or bile acid-binding
capacity./2"
Chitosan is relatively insoluble in water, but can be dissolved by dilute acids, which
would ma&e it a highly-viscous dietary fiber./5" ;uch fibers might inhibit the upta&e of
dietary lipids by increasing the thic&ness of the boundary layer of the intestinal lumen, which
has been observed in animal e*periments.
3aving very few acetyl groups, chitosan contains cationic groups. 'his may cause
chitosan to have bile acid-binding capacity, which causes mi*ed micelles to be entrapped or
disintegrated in the duodenum and ileum. /8" 'his would interrupt bile acid circulation,
causing reduced lipid absorption and increased sterol e*cretion, which has also been observed
in animal e*periments. 29"
,8 A..$i#ation+ o! C,ito+an
@a+t"-at"% T%"+"m"nt
Removed of 7etel lons
4locculent Coagulant
4rotein
$ye
#mino #cids
Foo1 In1(+t%
Colour ;tabitation
#nimal Iood #dditive
4emouned of $ye ;uspounded
;olid
M"1i#a$
Bandeges
Blood Cholestrol Control
Controlled Release of $rugs
;&in Bum
&iot"#,no$og
Cell Recovery
Chromatogrophy
Cell ,mmobili+ation
En+yme ,mmobili+ation
Ag%i#($t(%"
;eed Coating
Controlled #geochemical Release
Iertili+er
Co+m"ti#+
7oisturi+er
Iace ,3and and Body Creams
Beth Lotion
P($. an1 .a."%
;urface 'reatment
4hotographic 4aper.
M"m4%an"
4ermeability Control
Reverse (smesis
Chapter-? E*perimental ;ection
6. EBPERIMENTAL SECTION
6.1. MATERIALS:
Ta4$" no ;: Mat"%ia$+ (+"1 in in +it( g"$ation
S%. No. Mat"%ia$+ U+"1 G%a1" Man(!a#t(%"%
1 Gatiflo*acin 4harma #rrow chem,
2 4ola*amer !55 4harma #+ing pharma, hyd.
2 Carbapol 8). LR 7er& limited, 7umbai.
3 Chitosan 4harma #rrow chem.
6 Ben+al&onium Chloride LR Ranba*y Iine Chemicals Limited.
9 ;odium chloride LR 4oona Chemical Laboratory, 4une.
:
4otassium dihydrogen
4hosphate
LR
Ranba*y Iine Chemicals Limited,
Lew $elhi.
; ;odium hydro*ide LR 7er& limited, 7umbai.
< ;odium bicarbonate LR ;par& chem., Nolhapur.
1? Calcium chloride LR Lice chem, &ochin.
6.2. EFUIPMENTS
Ta4$" no <: E*(i.m"nt+ (+"1 in in +it( g"$ation
E*(i.m"nt Com.an
1 Electronic balance #fcoset, 7umbai.
2 7agnetic stirrer Remi E%uipments, 7umbai.
2 Cater $istillation Enit
Bhanu ;cientific ,ndustries, Co. 4vt. Ltd.,
Bangalore.
3 #utoclave 7.N. Laboratories, Bangalore.
6 I',R ;pectrophotometer 'hermo Licolet, E;#.
9 p3 meter Elico ,ndia ;ystronics, #hmedabad.
: Broo&field $H-!!!` Rheometer
Broo&field Engineering Laboratories ,nc.,
E;#.
; EH-His ;pectrophotometer ;himad+u, Yapan.
< 3ot air oven 4;7 ,ndustries, Bangalore.
6.2. METHODS:
6.2.1. P%"!o%m($ation +t(1i"+:
4reformulation testing is the first step in rational development of dosage forms of a
drug substance. ,t can be defined as investigation of physical and chemical properties of a
drug substance alone and when combined with e*icipients. 'he overall ob0ective of
preformulation testing is to generate information useful to the formulator in developing stable
and bio available dosage forms which can be mass produced. 2!"
18 I1"nti!i#ation:
a8 D"t"%mination o! m"$ting .oint:
7elting point of gatiflo*acin was determined by capillary method.
48 So$(4i$it:
;olubility is an important consideration in ophthalmic formulations as clarity
of the solution is an essential re%uirement. 'he solubility of gatiflo*acin, chloramphenecol
was detrmined in various solvents li&e distilled water, 9.! 7 hydrochloric acid, 9.27 sodium
hydrohide and some buffer solutions" in order to choose suitable solvent system.
#8 Com.ata4i$it +t(1i"+:
Compatability studies was carried out in order to establish, that there would be no
interaction between the drug and E*cipients eg: polymers" used in tne formulation. 'hese
studies were carried out by ir spectroscopy.
18 Stan1a%1 #a$i4%ation #(%)" o! gati!$o>a#in (+ing +im($at"1 t"a% !$(i1 7STF8:
;'I having the composition of sodium chloride 9./2 gm, sodium bicarbonate9.29
gm, calcium chloride 9.995 gm in !99 ml distilled water was prepared. #ccurately weighed
!99 mg gatiflo*acin was dissolved in minimum amount of 9.27 Lao3 solution and volume
was made up to !99 ml with ;'I to get the stoc& solution of !99 Dg1ml. Irom this stoc&
solution, ali%uots of 9.2, 9.., 9./, 9.5, !.9, !.2 and !.. mlwere withdrawn and diluted to !9 ml
with ;'I to get concentrations in the range of 2 to !. Dg1ml. 'he absorbance of these
solutions was measured at 25? nm by EH-His spectrophotometer.
"8 Stan1a%1 Ca$i4%ation C(%)" o! Gati!$o>a#in in Di+ti$$"1 @at"%:
#ccurately weighed !9 mg Gatiflo*acin was dissolved in distilled water to get the
stoc& solution of !99 Dg1ml. Irom this stoc& solution, ali%uots of 9.2, 9.., 9./, 9.5, !.9 and !.2
ml were withdrawn and volume was made up to !9 ml with distilled water to get
concentrations in the range of 2 to !2 Dg1ml. 'he absorbance of these solutions was measured
at 289 nm by EH-His spectrophotometer.
!8 Stan1a%1 #a$i4%ation #(%)" o! #,$o%am.,"ni#o$ (+ing +im($at"1 t"a% !$(i1 7STF8:
;'I having the composition of sodium chloride 9./2 gm, sodium bicarbonate9.29
gm, calcium chloride 9.995 gm in !99 ml distilled water was prepared. #ccurately weighed
!99 mg g chloramphenecol was dissolved in minimum amount of 9.27 Lao3 solution and
volume was made up to !99 ml with ;'I to get the stoc& solution of !99 Dg1ml. Irom this
stoc& solution, ali%uots of 9.2, 9.., 9./, 9.5, !.9 mlwere withdrawn and diluted to !9 ml with
;'I to get concentrations in the range of 2 to !9 Dg1ml. 'he absorbance of these solutions
was measured at 2.) nm by EH-His spectrophotometer.
g8 Stan1a%1 Ca$i4%ation C(%)" o! #,$o%am.,"ni#o$ in Di+ti$$"1 @at"%:
#ccurately weighed !9 mg chloramphenecol was dissolved in distilled water to get
the stoc& solution of !99 Dg1ml. Irom this stoc& solution, ali%uots of 9.2, 9.., 9./, 9.5, !.9 ml
were withdrawn and volume was made up to !9 ml with distilled water to get concentrations
in the range of 2 to !9 Dg1ml. 'he absorbance of these solutions was measured at 2.) nm by
EH-His spectrophotometer.
6.2.2. P%".a%ation o! In Sit( G"$$ing S+t"m o! Gati!$o>a#in:
a8 P%".a%ation o! P,o+.,at" 4(!!"% .H 9.;:5
?9 ml of potassium dihydrogen phosphate 9.2 7" and 22.. ml of sodiumhydro*ide
9.2 7" were mi*ed and volume was made up to 299 ml with water 22".
48 P%".a%ation o! in +it( g"$$ing ++t"m:5
$ifferent formulations were prepared with various ratios of pola*mer !55 and
carbopol 8). !5> to 2!> w1w": 9.9!> to 9.9.> w1w"
St". I: 'he method involved slow addition pola*mer !55 and methyl paraben were
solubili+ed in re%uired %uantity of cold distilled water.
St". II: Re%uired %uantities of carbopol 8). were &ept overnight for swelling.
St". III: re%uired %u%ntities of chitosan was dissolved in !?> acetic acid solution. 'he
polymer solution ta&en in a bea&er with continuous stirring magnetic stir" until uniform
solution obtained.
St". IV: #fter the mi*ture had been &ept at ambient 'emperature for 2. hrs. a smallamount
of trienthanolamine was added to ad0ust the 43 2.
St". V: #n appropriate amount of drug;olubili+ed in physiologically compatible solvent
such as sodium chloride was dissolved in phosphate buffer solution p3 /.5 with continues
stirring until uniform $rug solution obtained. 'hermo reversible gels were prepared using
cold techni%ue. 2)"
St". VI: $rug solution was added to these 4olymer solutions. 'he developed formulations
were filled in amber glass vials, closed with sterili+ed rubber closures and aluminium caps
and were sub0ected to crimping. 'he formulations in their final pac& were sub0ected to
terminal sterili+ation by autoclaving at !2!RC and !? psi for 29 mins. 2."
Ta4$" no 1?: Fo%m($ation 1"+ign !o% in +it( g"$$ing ++t"m+ o! gati!$o>a#in
Ing%"1i"nt+ F1 F2 F2 F3
Gatiflo*acin >w1v" 9.) 9.) 9.) 9.)
4ola*amer !55 > w1v" !5 !8 29 2!
Carbapol 8). >w1v" 9.9! 9.92 9.9) 9.9.
Chitosan >w1v" - - 9.9! 9.9?
7ethyl paeraben>w1v" 9.9! 9.9! 9.9! 9.9!
;odium chloride>w1v" 9.8 9.8 9.8 9.8
4hosphate buffer p3 /.5 ml" %s to !99 !99 !99 !99
6.2.2. P%".a%ation o! In Sit( G"$$ing S+t"m o! #,$o%am.,"ni#o$:
a8 P%".a%ation o! P,o+.,at" 4(!!"% .H 9.;:5
?9 ml of potassium dihydrogen phosphate 9.2 7" and 22.. ml of sodium hydro*ide
9.2 7" were mi*ed and volume was made up to 299 ml with water. 2?"
48 P%".a%ation o! in +it( g"$$ing ++t"m:5
$ifferent formulations were prepared with various ratios of pola*mer !55 and
carbopol 8). !5> to 2!> w1w": 9.9!> to 9.9.> w1w"
St". I: 'he method involved slow addition pola*mer !55 and methyl paraben were
solubili+ed in re%uired %uantity of cold distilled water.
St". II: Re%uired %uantities of carbopol 8). were &ept overnight for swelling.
St". III: Re%uired %u%ntities of chitosan was dissolved in !?> acetic acid solution. 'he
polymer solution ta&en in a bea&er with continuous stirring magnetic stir" until uniform
solution obtained.
St". IV: #fter the mi*ture had been &ept at ambient 'emperature for 2. hrs. a small amount
of trienthanolamine was added to ad0ust the 43 2.
St". V: #n appropriate amount of $rug ;olubili+ed in physiologically compatible solvent
such as sodium chloride was dissolved in phosphate buffer solution p3 /.5 with continues
stirring until uniform $rug solution obtained. 'hermo reversible gels were prepared using
cold techni%ue. 2/"
St". VI: $rug solution was added to these 4olymer solutions. 'he developed formulations
were filled in amber glass vials, closed with sterili+ed rubber closures and aluminium caps
and were sub0ected to crimping. 'he formulations in their final pac& were sub0ected to
terminal sterili+ation by autoclaving at !2!RC and !? psi for 29 mins.
Ta4$" no: 11 Fo%m($ation 1"+ign !o% in +it( g"$$ing ++t"m+ o! #,$o%am.,"ni#o$
Ing%"1i"nt+ F1 F2 F2 F3
Chloramphenicon>w1v" 9.2 9.2 9.2 9.2
4ola*amer !55 > w1v" !5 !8 29 2!
Carbapol 8). >w1v" 9.9! 9.92 9.9) 9.9.
Chitosan >w1v" - - 9.9! 9.9?
7ethyl paeraben>w1v" 9.9! 9.9! 9.9! 9.9!
;odium chloride>w1v" 9.8 9.8 9.8 9.8
4hosphate buffer p3 /.5 ml" %s to !99 !99 !99 !99
Chapter-/ Evalution ;tudies
9. EVALUTION STUDIES:
'he in situ gelling systems were evaluated for following parameters:
#ppearance
p3
$rug content
Gelation studies
;preadability
7easurement of Gel ;trengt
$etermination of mucoadhesive Iorce
Rheology
,n vitro release studies
;terility
,n vitro efficacy
Rabbit eye irritation
;tability studies
9.1. A.."a%an#":
Clarity of solution is one of the most important characteristic feature of ophthalmic
preparation. 'he appearance of the formulation was observed which included clarity and
colour of solution. 22"
9.2. D"t"%mination o! .H:
p3 is one of the most important factors involved in the formulation process. 'wo
areas of critical importance are the effects of p3 on solubility and stability. 'he p3 of
ophthalmic formulation should be such that the formulation will be stable at that p3 and at
the same time there would be no irritation to the patient upon administration of the
formulation. 'he p3 of the prepared formulations was chec&ed by using p3 meter.
9.2. D%(g Cont"nt:
Eniform distribution of active ingredient is important to achieve dose uniformity. 'he
drug content was determined by ta&ing !ml of the formulation and diluting it to !99 ml with
distilled water. #li%uot of ? ml was withdrawn and further diluted to 2? ml with distilled
water. Gatiflo*acin concentration was determined at 289 nm, Chloramphenicol concentration
was determined at 2.) nm by using EH-His spectrophotometer.
9.3. G"$ation St(1i"+:
'o mimic the situation where, in situ Gatiflo*acin system, upon ocular instillation, is
diluted with the available tear fluid and the gelation is induced by a limited supply of
electrolytes, the in situ gelling system was mi*ed with ;'I in the proportion 2?:2 application
volume 2? Dl, normal volume of tear fluid in the eye 2 Dl". Gelation was assessed by visual
e*amination. 25"
9.6. S.%"a1a4i$it:
Ior the determination of spreadability, e*cess of sample was applied in between two
glass slides and was compressed to uniform thic&ness by placing !999g weight for ? min.
weight ?9 g" was added to the pan. 'he time in which the upper glass slide moves over to the
lower plate was ta&en as measure of spreadability ;" 28".
;\ 7L1'
Chere, 7 \ weight tide to upper slide.
L \ length moved on the glass slide.
' \ time ta&en. 59"
9.9. M"a+(%"m"nt o! G"$ St%"ngt,:
# sample of ?9 gm of gel was placed in a !99 ml graduated cylinder and gelled in a
thermostat at )2
9
c. 'he apparatus for measuring gel strength weight of apparatus as shown in
figure !!, weighing 22 gm" was allowed to penetrate in occular gel. 'he gels strength, which
means the viscosity of the gels at physiological temperature, was determined by the time
seconds", the apparatus too& to sin& ?cm down through the prepared gel. 'he gels at
physiological temperature, was determined by the time seconds", the apparatus too& to sin&
?cm down through the prepared gel. 5!"
Fig. No. 16 G"$ +t%"ngt, 1"t"%mination 1")i#"
9.:. D"t"%mination o! m(#oa1,"+i)" Fo%#":
'he mucoadhesive force of all the optimi+ed batches was determined as follows, a
section of mucosa was cut from the Goat occular portion and instantly fi*ed with mucosal
side out onto each glass vial using rubber band. 52" 'he vial with occular mucosa was
connected to the balance in inverted position while first vial was placed on a height ad0ustable
pan. (ccular gel was added onto the nasal mucosa of first vial. Before applying the gel,
!?9DL of simulated tear solution was evenly spread on the surface of the test
memberane.'hen the height of second vial was so ad0usted that the mucosal surfaces of both
vials come in intimate contact. 'wo minutes time of contact was given. 'hen weight was &ept
rising in the pan until vials get detached. 52"
7ucoadhesive force was the minimum weight re%uired to detach two vials. 'he
chee& mucosa was changed for each measurement.
$etachment stress dynes1cm2" \ mg1#
Chere m is the weight added to the balance in grams-
g is the acceleration due to gravity ta&en as 859 cm1s2-
# is the area of tissue e*posed, i.e 2.!2 cm
2
.
#ll the above mentioned e*periments were carried out in triplicates.
Fig. No. 19 M(#oa1,"+i)" Fo%#" m"a+(%ing 1")i#"
7A8 Mo1i!i"1 &a$an#". 7&8 @"ig,t+ 7C8 G$a++ )ia$ 7D8 o#($a% in +it( g"$ 7"8 o#($a%
m"m4%an" 7F8 H"ig,t a1'(+ta4$" .an.
9.;. R,"o$og:
Hiscosity of instilled formulation is an important factor in determining residencetime
of drug in the eye. 'he viscosity determination of prepared formulations was carried out using
Broo&filed $H-!!!` Rheometer with spindle LH-). 'he prepared sol was allowed to gel in
the ;'I and then viscosity was measured. Hiscosity of samples was measured at different
angular velocities. # typical run comprised changing angular velocity from !9 to !99 rpm
with e%ual wait for each rpm. 'he hierarchy of angular velocity was reversed !99 to!9 rpm"
with similar wait. 'he average of two readings were usedto calculate the viscocity. 5)"
9.<. In )it%o %"$"a+" +t(1i"+:
'he in vitro release of gatiflo*acin, chloramphenicol from the formulation was
studied through goat ocular membrane using modified apparatus. 'he diffusion medium used
was ;'I, freshly prepared p3 2..". (cular membrane previously soa&ed overnight in the
dissolution medium was tied to one end of specially designed glass cylinder opened at both
cylinders". ! ml of formulation e%ual to ) mg of gatiflo*acin" was accuratetly placed in to
this assembly. 5)" 'he cylinder was attached to a stand and suspended in ?9 ml of diffusion
medium maintained at )2F!
9
c so that the membrane 0ust touched the receptor medium
surface. 'he diffusion medium was stirred at low speed using magnetic stirrer. #li%uots, each
of ? ml volume were with drawn at hourly intervals and replaced by an e%ual volume of
receptor medium. 'he ali%uots were suitably diluted with the receptor medium and anlysed by
EH visible spectrophotometry at 289 nm for gatiflo*acin formulations and 2.. nm for
chloramphenicol formulations. 5."
9.1?. St"%i$it:
#ll ophthalmic preparations should be sterile therefore the test for sterility is very
important evaluation parameter. 'he sterility test was performed according to ,ndian
4harmacopoeia. $irect inoculation method was used. 2 ml of li%uid from test container
wasremoved with a sterile pipette or with a sterile syringe or a needle. 'he test li%uid was
aseptically transferred to fluid thioglycolate medium 29 ml" and soyabean-casein digest
medium 29 ml" separately. 'he li%uid was mi*ed with the media. 'he inoculated media were
incubated for not less than !. days at )9RC to )?RC in the case of fluid thioglycolate medium
and 29RC to 2?RC in the case of soyabean-casein digest medium. 5?"
9.11. In )it%o E!!i#a#:
#ntimicrobial efficacy studies were carried out to ascertain the biological activity of
sol-to-gel systems against micro-organisms. 'his was determined by agar diffusion test
employing Jcup plate techni%ueK. ;terile solution of Gatiflo*acin mar&eted product G#'E
EaE $R(4; manufactured by #0anta 4harma Ltd., B.Lo.74((23E, 7fd. 7ay 29!9, E*p.
#pr 29!2" as a standard was used and the developed formulations test solutions" were poured
into cups bored into sterile 7uller 3inton #gar 73#" previously seeded with test organisms
4seudomonas aeruginosa and ;taphylococcus aureus". #fter allowing diffusion of solutions
for 2 hr, the plates were incubated for 2. hrs at )2RC. 'he +one of inhibition Q(," measured
around each cup was compared with that of standard. Both positive and negative controls
were maintained throughout the study. 5/, 52"
9.12. Ra44it "" i%%itation:
(cular irritation studies were performed on 2 male rabbits each weighing !-2 &g. 'he
sterile formulations were instilled twice a day for a period of 2! days and the rabbits were
observed periodically for redness, swelling and watering of the eye. 55"
Chapter-2 Results #nd $iscussion
:. RESULSTS AND DISCUSSION
:.1. P%"!o%m($ation St(1i"+:
I. I1"nti!i#ation:
a8 D"t"%mination o! M"$ting Point:
7elting point of Gatiflo*acin was found to be in the range of !5!RC to !5)RC as reported in
literature, thus indicating purity of the drug sample as any impurity if present will cause
variations in the melting point of a given drug substance.
7elting point of chloramphenecol was found to be 88RC as reported in literature,
thus indicating purity of the drug sample as any impurity if present will cause variations in the
melting point of a given drug substance
48 So$(4i$it:
Gatiflo*acin was found to be freely soluble in 9.27 sodium hydro*ide, 9.!L
hydrochloric acid and very slightly soluble in water. 'he solubility of Gatiflo*acin was found
to be p3 dependent.
Chloramphenicol was found to be freely soluble in ethanol, methanol, chloroform
9.27 sodium hydro*ide, 9.!L hydrochloric acid and poorly soluble in water. 'he solubility
of chloramphenecol was found to be p3 dependent.
II. Com.ati4i$it:
,nfra red spectra of pure drug Gatiflo*acin, carbapol8)., pola*amer !55 and
combination of drug with the polymers were obtained which are shown in spectra no.!, 2, ), .
and ? respectively. #ll the characteristic pea&s of Gatiflo*acin were present in spectrum no.2
thus indicating compatibility between drug and the polymers. 'he spectrum confirmed that
there is no significant change in chemical integrity of the drug.
$;C studies of pure drug chloramphenicol, carbapol8)., pola*amer !55 and
combination of drug with the polymers were obtained which are shown ingraph no./, 2, 5, 8
and !9 respectively. #ll the characteristic pea&s of chloramphenicol were present in spectrum
no.5 thus indicating compatibility between drug and the polymers. 'he spectrum confirmed
that there is no significant change in chemical integrity of the drug.
Fig. No. 1: FTIR +."#t%a o! .o$o>am"% 1;;
Fig. No. 1; FTIR +."#t%a o! .(%" 1%(g
Fig. No. 1< FTIR +."#t%a o! #,ito+an
Fig. No. 2? FTIR +."#t%a o! #a%4a.o$ <23
Fig. No. 21 FTIR +."#t%a o! !o%m($ation
Fig. No. 22 DSC +."#t%a o! .o$o>am"% 1;;
Fig. No. 22 DSC +."#t%a o! .(%" 1%(g
Fig. No. 23 DSC +."#t%a o! #,ito+an
Fig. No. 26 DSC +."#t%a o! #a%4a.o$
Fig. No. 29 DSC +."#t%a o! !o%m($ation
Ta4$" no: 12 FTIR +."#t%a o! P(%" 1%(g
St%"t#,ing R".o%t"1 .(%"1%(g
P%o#(%"1 .(%" 1%(g
)a$("+
Fo%m($ation
Ca%4o>$at" 2313 2312.1; 2323.:9
S"#on1a% amin" 166? 163<.39 1661.:3
Con'(gat"1 D""ton" 192: 1922.21 1922.2;
A$Dan" CH
2
2<9? 2<:6.19 2<:1.62
A%omati# CH <??59:?
9;;.22
:29.62
::2.?:
;3:.12
;<1.3;
9;;.2<
:26.62
;21.;2
;32.31
;<2.22
Ta4$" no: 12 FTIR +."#t%a o! Po$a>am"% 1;;
St%"t#,ing R".o%t"1 .(%"1%(g
P%o#(%"1 .(%" 1%(g
)a$("+
Fo%m($ation
A$i.,ati#
C5H +t%"t#,ing
2;<1 2;;;.2; 2;;:.;2
P$an"
O5H 4on1
1232 1232.3; 1922.2;
C5H +t%"t#, 1111 1112.21 1112.63
Ta4$" no: 13 FTIR +."#t%a o! #,ito+an
St%"t#,ing R".o%t"1 .(%"1%(g
P%o#(%"1 .(%" 1%(g
)a$("+
Fo%m($ation
5NH
2
233< 2326.91 2323.:<
5OH 196? 1923.?? 1922.2;
Ta4$" no: 16 FTIR +."#t%a o! #a%4a.o$ <23
St%"t#,ing R".o%t"1 .(%"1%(g
P%o#(%"1 .(%" 1%(g
)a$("+
Fo%m($ation
C5H 2<2? 2<21.31 2<:1.62
#a%4o>$at" amin" 1:2? 1:?<.29 5
O5H 4"n1 111? 1112.6: 1113.?;
Stan1a%1 Ca$i4%ation C(%)" o! Gati!$o>a#in in STF:
'he absorbance values are shown in 'able !/. 'he Beers range of Gatiflo*acin in
;'I was found to be 2 to !. Dg1ml at 25? nm by EH-His spectrophotometer. Graph !! shows
the plot of absorbance Hs. concentration.
Ta4$" no: 19 Stan1a%1 Ca$i4%ation C(%)" o! Gati!$o>a#in in STF
Fig. No. 2: Stan1a%1 Ca$i4%ation C(%)" o! Gati!$o>a#in in STF
Stan1a%1 Ca$i4%ation C(%)" o! Gati!$o>a#in in 1i+ti$$"1 -at"%:
'he absorbance values are shown in 'able !2. 'he Beers range of Gatiflo*acin in
;'I was found to be 2 to !2 Dg1ml at 289 nm by EH-His spectrophotometer. Graph !2 shows
Ta4$": No 1: Stan1a%1 #(%)" o! Gati!$o>a#in at 2<? nm in 1i+ti$$"1 -at"%
S$.no. Con#"nt%ation m#g=m$ A4+o%4an#"
!. 9 9
2. 2 9.!)2
). . 9.22.
.. / 9..!)
?. 5 9.?)!
/. !9 9./.5
2. !2 9.522
5. !. 9.8..
!. 9 9
2. 2 9.!/5
). . 9.)2.
.. / 9..59
?. 5 9./2/
/. !9 9.5).
2. !2 9.822
Fig. No. 2; Stan1a%1 Ca$i4%ation C(%)" o! Gati!$o>a#in in 1i+ti$$"1 -at"%
Stan1a%1 Ca$i4%ation C(%)" o! #,$o%am.,"ni#o$ in STF:
'he absorbance values are shown in 'able !5. 'he Beers range of chloramphenicol in
;'I was found to be 2 to !9 Dg1ml at 2.) nm by EH-His spectrophotometer. Graph !) shows
Ta4$". No: 1; Stan1a%1 #(%)" o! #,$o%am.,"ni#o$ at 232 nm in STF
Fig. No. 2< Stan1a%1 #(%)" o! #,$o%am.,"ni#o$ at 232 nm in STF
Stan1a%1 Ca$i4%ation C(%)" o! #,$o%am.,"ni#o$ in 1i+ti$$"1 -at"%:
'he absorbance values are shown in 'able !8. 'he Beers range of chloramphenicol in
distilled water was found to be 2 to !9 Dg1ml at 2.. nm by EH-His spectrophotometer. Graph
!. shows the plot of absorbance Hs.concentration.
Ta4$" No. 1< Stan1a%1 #(%)" o! Gati!$o>a#in at 233 nm in 1i+ti$$"1 -at"%
!. 9 9
2. 2 9.9)2
). . 9.958
.. / 9.988
?. 5 9.!8/
/. !9 9.2.)
!. 9 9
2. 2 9.9?)
). . 9.!98
.. / 9.!?5
?. 5 9.2!)
/. !9 9.2/2
Fig. No. 2? Stan1a%1 Ca$i4%ation C(%)" o! #,$o%am.,"ni#o$ in 1i+ti$$"1 -at"%
E)a$(ation +t(1i"+:
1. A.."a%an#":
Clarity of all formulations was found to be satisfactory. 'he formulations were light
yello in colour. 'erminal sterili+ation with autoclaving had no effect on the physic chemical
properties of the formulations. 'he ha+iness that was observed after autoclaving was found
disappears and the original clarity was regained after over night standing.
2. .H 1"t"%mination:
p3 values for all the formulations are given in table 29, 2!. 'he p3 was with in
acceptable range and hence would not cause any irritation upon administration of the
formulation.
Ta4$" No.2? .H an1 1%(g #ont"nt o! .%".a%"1 in+it( g"$$ing ++t"m o! Gati!$o>a#in
Fo%m($ation .H D%(g #ont"nt G
I! /.5 82..)
I2 /.5 !9!./5
I) /.5 !9)...
I. /.5 !92.)!
Ta4$" No. 21 .H an1 1%(g #ont"nt o! .%".a%"1 in+it( g"$$ing ++t"m o! C,$o%am.,"ni#o$
Fo%m($ation .H D%(g #ont"nt G
I! /.5 8/.?2
I2 /.5 !9!.25
I) /.5 !99.82
I. /.5 !9!.)52
2. D%(g Cont"nt:
'able 29, 2! shows the result of percent drug content for all the formulations. 'he
drug content was found to be in acceptable range for all the formulations. > $rug content in
all eight formulations was in the range 8/.?2 G !9)...> indicating uniform distribution of
drug.
3. G"$ation +t(1i"+:
'he two main prere%uisites of gelling system are viscosity and gelling capacity
speed and e*tent of gelation". 'he formulation should have an optimum viscosity, which will
allow its easy instillation into the eye as a li%uid drops", which will then undergo rapid sol to
gel transition due tochange in temparature. 7oreover, to facilitate sustained release of drug to
the ocular tissue, the in situ formed gel should preserve its integrity without dissolving or
eroding for a prolonged period of time. #ll the formulations gelled instantaneously less than
a minute" on contact with ;'I in the body temparature. By visual inspection, the formulations
formed a translucent matri* on addition to the ;'I. 'he gelation may be due change in
temperature.
6. G"$ St%"ngt,:
#ll three formulations I2, I) and I." e*hibited good gel strength. 'his may be due
to increase concentration of chitosan and carbapol 8). along with constant ratio ofpola*amer
!55. ;econd method also showed good gel strength by comparing with I!.
9. M(#oa1,"+i)" Fo%#":
'he mucoadhesive force is an important physicochemical parameters for prolonging
occularretention time and there by better therapeutic effects. $etachment stress of I. was
found to more in comparison with I! formulation. 'he formulation f2, I), I. showed more
mucoadhesive force than I!. 'his may be due to increased concentration of Chitosan and
carbapol along with pola*amer !55 in the formulation.
:. S.%"a1a4i$it:
I. showed good spreadability as copared with the any other formulatin. Comparing
I) and I. showed good spreadability by comparing with I! and I2. 'he formulation from
thermoreverse gelation showed good spreadability comparing with other formulations.
Ta4$" No. 22 S.%"a1a4i$itA g"$+t%"ngt,A m(#oa1,"+i)" !o%#" o! .%".a%"1 in+it( g"$$ing
++t"m o! gati!$o>a#in
Fo%m($ation
M(#oa1,"+i)"
!o%#"71n"+=#m28
G"$
+t%"ngt,7+"#8
S.%"a1a4i$it
gm+=+"#
I! !2/!2 88 22.9
I2 !/5)2 !!9 2/.?
I) !2?22 !!) 25.2
I. !22/9 !!? 28.?
Ta4$" No. 22 +.%"a1a4i$itA g"$+t%"ngt,A m(#oa1,"+i)" !o%#" o! .%".a%"1 in+it( g"$$ing
++t"m o! #,$o%am.,"ni#o$
Fo%m($ation
M(#oa1,"+i)"
!o%#"71n"+=#m28
G"$
+t%"ngt,7+"#8
S.%"a1a4i$it
gm+=+"#
I! !)?25 !99 22.2
I2 !22.. !98 2?.?
I) !5/8. !!2 )9.2
I. !58/9 !!8 )!.2
;. R,"o$ogi#a$ St(1i"+:
'able 2. shows the viscosity values obtained for all the formulations of gatiflo*acin
and table 2? shows the viscosity of chloramphenicol using Broo&field $H-!!!` rheomoter.
'he formulations e*hibited pseudoplastic rheology, as evidenced by shear thinning and an
increase in shear stress with increased angular velocity. 'he viscosity was directly dependent
on the polymeric content of the formulation. 'he viscosity increased with increasing
concentration of carbapol and chitosan. I. showed the ma*imum viscosity of .2 cps at 29
rpm whereas the minimum viscosity at 29 rpm was shown by I!. 'his indicated that addition
of carbapol 8). and chitosan led to increase in viscosity
Ta4$" No. 23 Vi+#o+it o! .%".a%"1 in+it( g"$$ing ++t"m o! gati!$o>a#in
Ang($a%
)"$o#it7%.m8
Vi+#o+it
F1 F2 F2 F3
!9 /2.2 2! 82 88
29 28.) )2 )5 .2
)9 2..2 2? 2. 25.2
.9 !5.8 !8.? !8.? 2..2
?9 !).2 !8.? !2.2 22.2
/9 !9.2 !..2 !).. !5..
29 5.? !2.5 !2 !2.)
59 5 8.? 8.? !...
89 2.. 5.!. 5.2 !..!
!99 /.? 2./! 2.5 !)..
Ta4$" No. 26 )i+#o+it o! .%".a%"1 in+it( g"$$ing ++t"m o! #,$o%am.,"n"#o$
Ang($a%
)"$o#it7%.m8
Vi+#o+it
F1 F2 F2 F3
!9 /9.2 /5 8. 88
29 25 )!.8 .2 .8
)9 2).. 2..5 25 28.2
.9 !2.. !8.5 29.? 2/.2
?9 !2./ !5.! !8.2 2..5
/9 8.!2 !).? !?.. !8./
29 2.2 !2./ !)./ !5.)
59 2 8.8 !9.? !?..
89 /.5 5.2. 8.5 !..!
!99 /.2 2./ 5.5 !)..
<. In )it%o %"$"a+" +t(1i"+:
'he in vitro release &inetics was carried out for all formulations using ;'I as the
diffusion medium. 'he data of these studies are presented in tables
,t was found that drug release was for the formulations I!-I. respectively after / hr.
these values indicates that I. showed better sustaining effect among all formulations. 'his
may due to the higher concentrtations of carbapol, chitosan and popla*amer in I.. Iigure
shows the plot of in vitro release studies for all the formulations.
#ll formulations of chloramphenicol shows sustained release than gatiflo*acin
formulations with in / hr. this may due to the low permiability of the chloramphenicol.
'he drug release pattern obtained for the gelled samples is characteristic for
hydrophilic matrices. 'he initial fact the release of gatiflo*acin can be e*plained by the fact
that pola*amer eye drops are formulated in water and hence the polymer was completely
hydrated. Chen they e*posed to body temperature with ;'I and gelation occrs, a pre
hydrated matri* is formed in which hydration and water penetration no longer limit drug
release leading to an apparent diffusion-controlled release.
'he results obtained in vitro release studies were plotted in different modes of
data treatment as follows.
Cumulative percent drug released Hs. time Qero order rate &inetics".
Log cumulative percent drug retained Hs. time Iirst order &inetics".
Log cumulative percent drug released Hs. log time 4eppas e*ponential e%uation".
Cumulative percent released Hs. s%uare root of time 3iguchiAs classical diffusion
e%uation".
'he &inetic values obtained for different formulations are indicated in 'ables 2/
and 22.
'he regression values for formulations of gatiflo*acin I! to I. of first order plots
were found to be 9.8/!, 9.82., 9.8/! and 9.822 respectively.
'hese results indicated that formulations I! followed higuchi order &inetics whereas
I2, I) and I) followed +ero order. Graphical representation of +ero order and first order plots
is shown in Iigures . and ?.
'he regression values for formulations of chloramphenicol I! to I. of first order
plots were found to be 9.8/5, 9.82?, 9.825 and 9.82/ respectively.
Ta4$" No. 29 In )it%o G 1%(g %"$"a+" o! .%".a%"1 in+it( g"$$ing ++t"m o! gati!$o>a#in
Tim" 7,%8 F1 F2 F2 F3
9 9 9 9 9
9.? !2.?9. !!.85/ 8.)!. 2.2.)
! 25.888 22.9?5 2..292 2!.?/2
!.? .!.5)) .9.?92 )/./?2 )2.!/2
2 ?).!8/ ?2...8 .5./5. .).8?5
) /!.8.5 /9.5!2 ?/./2/ ?!.)2!
. //.228 /?.2?9 /!../8 ?/.298
? 2!.8)! 29.295 //..92 /9.))2
/ 2?..)9 2...5 /8.22 /).85
Fig. No. 21 In )it%o G 1%(g %"$"a+" o! .%".a%"1 in+it( g"$$ing ++t"m o!
Gati!$o>a#in
Ta4$" No. 29 In )it%o G 1%(g %"$"a+" o! .%".a%"1 in+it( g"$$ing ++t"m o!
C,$o%am.,"ni#o$
Tim" F1 F2 F2 F3
9 9 9 9 9
9.? !9.25 8.2/ 5.)25 2.2)
! 2?.85 2..22 22..) 2!.2)
!.? )?.52 )/.2/ )2./? )!./?
2 .2.25 .5.2. .?.!2 .)./)
) ??./8 ?/.2. ?).9/ ?!.5)
. /9.?8 /!.222 ?2.!/ ?/.)2
? /?..5 //.)2 /2.9. /9..2
/ 2!.22 /8.?/ //.28 /).22
Fig. No. 22 In )it%o G 1%(g %"$"a+" o! .%".a%"1 in+it( g"$$ing ++t"m o!
C,$o%am.,"ni#o$
Ta4$" no.2; R"$"a+" Din"ti# 1ata o! !o%m($ation F1 o! #,$o%am.,"n"#o$
Tim"
Log
Tim"
S*(a%"
%oot
Tim"
G 1%(g
%"$"a+"
Log G
1%(g
%"$"a+"
C(mm($ati)" G
1%(g %"$"a+"
Log G
C(m
1%(g
%"$"a+"
C(4"t,%ot
D%(g
%"maining
9 9 9 9 9 9 9 ../.!
9.? 9 9.292 !9.25 9.)!) !9.25 !.92 ...2/
! 9 ! 2?.85 9.2!?2 !2.))/ !.98!2 ..!85
!.? 9.!2/ !.22. )?.52 9.5??2 .).922 !./).2 ..99)
2 9.)9! !..!.2 .2.25 9.859) /2.!8/ !.28)5 ).2)25
) 9..22 !.2)2 ??./8 !.9./5 28.//2 !.89!) ).?)5
. 9./92 2 /9.?8 !.95). 8?.2 !.8598 )..9)!
? 9./85 2.2)/ /?..5 !.!!2! !!2.295 2.9?! ).2?/
/ 9.225 2...8 2!.22 !.!?)/ !)!.?. 2.!!8! ).9/.
Fig. No. 22 E"%o o%1"% %"$"a+" o! F1 o! #,$o%am.,"ni#o$
Fig. No. 23 Fi%+t o%1"% %"$"a+" o! F1 o! #,$o%am.,"ni#o$
Fig. No. 26 Hig(#,i %"$"a+" o! F1 o! #,$o%am.,"ni#o$
Fig. No. 29 Hi>+on #%o-"$$ %"$"a+" o! F1 o! #,$o%am.,"ni#o$
Fig. No. 2: 0o%+"m""% %"$"a+" o! F1 o! #,$o%am.,"ni#o$

Ta4$" no. 2< R"$"a+" Din"ti# 1ata o! !o%m($ation F2 o! #,$o%am.,"ni#o$
Tim" $ogT
S*(a%"
%oot T
G D%(g
R"$"a+"
LogGD%(g
R"$"a+"
GC(mm($ati)"
D%(g %"$"a+"
$ogG
C(mm($ati)"
D%(g R"$"a+"
C(4"t,%oot
D%(g%"maining
9 9 9 9 9 9 9 ../.!
9.? 9 9.292 8.2/ 9.8// 8.2/ 9.8/8 ..222
! 9 ! 2..22 !.)8 2/.?. !.?.5 ).85.
!.? 9.!2/ !.22. )/.2/ !.?/ .).?? !./)/ ).22/
2 9.)9! !..!.2 .5.2. !./5) /2.)5 !.282 ).295
) 9..22 !.2)2 ?/.2. !.2? 59.?2 !.89. ).?!
. 9./92 2 /!.22 !.25 8/.)2 !.85. ).)5.
? 9./85 2.2)/ //.)2 !.522 !!)./ 2.9?? ).22
/ 9.225 2...8 /8.?/ !.5. !)9.22 2.!!) ).!2
Fig. No. 2; E"%o o%1"% %"$"a+" o! F2 o! #,$o%am.,"ni#o$
Fig. No. 2< Fi%+t o%1"% %"$"a+" o! F2 o! #,$o%am.,"ni#o$
Fig. No. 3? Hig(#,i %"$"a+" o! F2 o! #,$o%am.,"ni#o$
Fig. No. 31 0o%+"m""% %"$"a+" o! F2 o! #,$o%am.,"ni#o$
Ta4$" no.2? %"$"a+" Din"ti# 1ata o! !o%m($ation F2 o! #,$o%am.,"n"#o$
Tim" $ogT
S*(a%"
%oot T
G D%(g
R"$"a+"
Log G
D%(g
R"$"a+"
GC(mm($ati)"
D%(g R"$"a+"
$ogG
C(mm($ati)"D%(g
R"$"a+"
C(4"t,%oot
D%(g
%"maining
9 9 9 9 9 9 9 ../.!
9.? 9 9.292 5.)25 9.82) 5.)25 9.82) ..?95
! 9 ! 22..) !.)? 2..!9? !.)52 ..2/.
!.? 9.!2/ !.22. )2./? !.?!) )5.5!! !.?55 ..9/5
2 9.)9! !..!.2 .?.!2 !./?. ?2.5!! !.2/2 ).5
) 9..22 !.2)2 ?).9/ !.22. 2..2. !.52) )./92
. 9./92 2 ?2.!/ !.2?2 58..? !.8?! )..88
? 9./85 2.2)/ /2.9. !.282 !9?.2 2.92 ).)/
/ 9.225 2...8 //.28 !.52! !22.) 2.952 ).2)
Fig. No. 32 E"%o o%1"% %"$"a+" o! F2 o! #,$o%am.,"ni#o$
Fig. No. 33 Fi%+t o%1"% %"$"a+" o! F2 o! #,$o%am.,"ni#o$
Fig. No. 36 Hig(#,i %"$"a+" o! F2 o! #,$o%am.,"ni#o$
Fig. No. 3: Hi>+on #%o-"$$ %"$"a+" o! F2 o! #,$o%am.,"ni#o$
Ta4$" no. 21 R"$"a+" Din"ti# 1ata o! !o%m($ation F3 o! #,$o%am.,"n"#o$
Tim" $ogT
S*(a%"
%oot T
G D%(g
R"$"a+"
LogG
D%(g
R"$"a+"
GC(mm($ati)"
D%(g R"$"a+"
$ogG
C(mm($ati)"
D%(g R"$"a+"
C(4"t,%oot
D%(g
%"maining
9 9 9 9 9 9 9 ../.!
9.? 9 9.292 2.2) 9.5?8 2.2) 9.5?8 ..?2?
! 9 ! 2!.2) !.))2 2).!2/ !.)/? ..222
!.? 9.!2/ !.22. )!./? !.? )2...2 !.?2) ..95
2 9.)9! !..!.2 .)./) !./)8 ??.2?2 !.2./ ).5).
) 9..22 !.2)2 ?!.5) !.2!. 22./2 !.5/! )./)
. 9./92 2 ?/.)2 !.2? 52..?/ !.8.! ).?2!
? 9./85 2.2)/ /9..2 !.25! !92.52 2.9! )..92
/ 9.225 2...8 /).22 !.5 !!2./?2 2.92 ).)95
Fig. No. 3; E"%o o%1"% %"$"a+" o! F3 o! #,$o%am.,"ni#o$
Fig. No. 3< Fi%+t o%1"% %"$"a+" o! F3 o! #,$o%am.,"ni#o$
Fig. No. 6? Hig(#,i %"$"a+" o! F3 o! #,$o%am.,"ni#o$
Ta4$" no.22 R"$"a+" Din"ti# 1ata o! !o%m($ation F1 o! gati!$o>a#in
Tim" $ogT
S*(a%"
%oot T
G D%(g
R"$"a+"
LogG
D%(g
R"$"a+"
GC(mm($ati)"
D%(g R"$"a+"
$ogG
C(mm($ati)"
D%(g R"$"a+"
C(4"t,%oot
D%(g
%"maining
! 9 ! 25.888 !../2 25.888 !..85 ..!.
!.? 9.!2/ !.22. .!.5)) !./2 .!.5) !.2 ).52.
2 9.)9! !..!.2 ?).!8/ !.22? ?).!8 !.5.. )./9)
) 9..22 !.2)2 /!.8.5 !.282 /!.8. !.8? ).)/)
. 9./92 2 //.228 !.52. //.22 2.92/ ).2!/
? 9./85 2.2)/ 2!.8)! !.5?/ 2!.8) 2.982 ).9)8
/ 9.225 2...8 2?..) !.522 2?..) 2.!?. 2.892
Fig. No. 62 E"%o o%1"% %"$"a+" o! F1 o! gati!$o>a#in
Fig. No. 63 Fi%+t o%1"% %"$"a+" o! F1 o! gati!$o>a#in
Fig. No. 66 Hig(#,i %"$"a+" o! F1 o! gati!$o>a#in
Fig. No. 69 Hi>+on #%o-"$$ %"$"a+" o! F1 o! gati!$o>a#in
Fig. No. 6: 0o%+"m""% %"$"a+" o! F1 o! gati!$o>a#in
Ta4$" no.22 R"$"a+" Din"ti# 1ata o! !o%m($ation F2 o! Gati!$o>a#in
Tim" $ogT
+*(a%"
%oot T
GD%(g
R"$"a+"
LogG
D%(g
R"$"a+"
GC(mm($ati)"
D%(g R"$"a+"
$ogG
C(mm($ati)"D%(g
R"$"a+"
C(4"t,%oot
D%(g
%"maining
9 9 9 9 9 9 9 ../.!
9.? 9 9.292 !!.85 !.925 !!.85 !.925 ....5
! 9 ! 22.9? !..)2 28... !../5 ..!25
!.? 9.!2/ !.22. .9.? !./92 .5.) !./5) ).89)
2 9.)9! !..!.2 ?2... !.2!8 /5.). !.5). )./22
) 9..22 !.2)2 /9.5!2 !.25) 52.!8 !.8. ).)8/
. 9./92 2 /?.2? !.5!2 !9..!8 2.9!2 ).2.2
? 9./85 2.2)/ 29.2 !.5./ !2!.58 2.95? ).988
/ 9.225 2...8 2...5 !.52 !.9.2! 2.!./ 2.8..
Fig. No. 6; E"%o o%1"% %"$"a+" o! F2 o! gati!$o>a#in
Fig. No. 6< Fi%+t o%1"% %"$"a+" o! F2 o! gati!$o>a#in
Fig. No. +9? Hig(#,i %"$"a+" o! F2 o! gati!$o>a#in
Fig. No. 91 0o%+"m""% %"$"a+" o! F2 o! gati!$o>a#in
Ta4$" no.23 %"$"a+" Din"ti# 1ata o! !o%m($ation F2 o! gati!$o>a#in
Tim" $ogT
S*(a%"
%oot T
GD%(g
R"$"a+"
LogG
D%(g
R"$"a+"
GC(mm($ati)"
D%(g R"$"a+"
$ogG
C(mm($ati)"
D%(g R"$"a+"
C(4"t,%oot
D%(g
%"maining
9 9 9 9 9 9 9 ../.!
9.? 9 9.292 8.)!. 9.8/8 9.8). 9.8/8 ...82
! 9 ! 2..292 !.)5) )?.)25 !.?.5 ..2)2
!.? 9.!2/ !.22. )/./? !.?/ .).)? !./)/ ).85/
2 9.)9! !..!.2 .5./5 !./52 /2.2!/ !.282 ).2!/
) 9..22 !.2)2 ?/./2 !.2?) 59.)2/ !.89. ).?!)
. 9./92 2 /!../ !.255 8/..5? !.85. ).)22
? 9./85 2.2)/ //.. !.522 !!).2 2.9? ).22/
/ 9.225 2...8 /8.22 !.5. !28.5 2.!!) ).!)
Fig. No. 93 Fi%+t o%1"% %"$"a+" o! F2 o! gati!$o>a#in
Fig. No. 96 Hig(#,i %"$"a+" o! F2 o! gati!$o>a#in
Fig. No. 99 Hi>+on #%o-"$$ %"$"a+" o! F2 o! gati!$o>a#in
Fig. No. 9: 0o%+"m""% %"$"a+" o! F2 o! gati!$o>a#in
Ta4$" no. 26 %"$"a+" Din"ti# 1ata o! !o%m($ation F3 o! gati!$o>a#in
Tim" $ogT
S*(a%"
%oot T
GD%(g
R"$"a+"
LogG
D%(g
R"$"a+"
GC(mm($ati)"
D%(g R"$"a+"
$ogG
C(mm($ati)"
D%(g R"$"a+"
C(4"t,%oot
D%(g
%"maining
9 9 9 9 9 9 9 ../.!
9.? 9 9.292 2.2.) 9.555 2.2.) 9.555 ..?!5
! 9 ! 2!.?/2 !.))) 2).!! !.)/) ..25
!.? 9.!2/ !.22. )2.!/2 !.?92 )5.92 !.?5 ..9/
2 9.)9! !..!.2 .).8?5 !./.2 ?/.2. !.2? ).52/
) 9..22 !.2)2 ?!.)2! !.2! 22.)8 !.5? )./?!
. 9./92 2 ?/.298 !.2.8 52.?? !.8. ).?2.
? 9./85 2.2)/ /9.))2 !.25 !92.898 2.9!2 )..!
/ 9.225 2...8 /).85 !.59/ !!5./2 2.92 ).)92
Fig. No. 9; E"%o o%1"% %"$"a+" o! F3 o! gati!$o>a#in
Fig. No. 9< Fi%+t o%1"% %"$"a+" o! F3 o! gati!$o>a#in
Fig. No. :? Hig(#,i %"$"a+" o! F3 o! gati!$o>a#in
Fig. No. :1 0o%+"m""% %"$"a+" o! F3 o! gati!$o>a#in
Ta4$" no. 29 Gati!$o>a#in %"$"a+" Din"ti#+
Ta4$" no. 2: C,$o%am.,"ni#a$ R"$"a+" Din"ti#+
S.no E"%o o%1"% Fi%+t o%1"% Hig(#,i 0o%+m""% Hi>+on C%o-"$$
R
2
m R
2
m R
2
m R
2
m R
2
m
!
9.8/5 22.?!* 9./28 9.22) 9.822 ?5./5* 9./28 !.9?/ 9.8)2 9.2?/
2
9.82? 2!.58 9./!/ 9.2?/ 9.8?2 ?2./9 9.?!2 !.)52 9.52) 9.229
)
9.825 29.?2 9./?) 9.2/! 9.8.8 ?).8? 9.?)? !.)8! 9.82/ 9.2)?
.
9.82/ !8.85 9.//9 9.2/) 9.8.5 ?2..) 9.?.. !..92 9.8!/ 9.22?
S.no E"%o o%1"% Fi%+t o%1"% Hig(#,i 0o%+m""% Hi>+on C%o-"$$
R
2
m R
2
m R
2
m R
2
m R
2
m
! 9.5?2 !!.8? 9./!2 9.2?2 9.8/! )).28 9..28 !.))8 9.82)
9.229*
2 9.82. 2).)? 9./29 9.2?5 9.8?2 /!./! 9..58 !.)?9 9.82)
9.228
) 9.8/! 2!.8/* 9./!? 9.2?? 9.8.? ?5.99 9.?!8 !.)5/ 9.8!5
9.2?.
. 9.822 29.9) 9./?2 9.2/! 9.8.5 ?2.?? 9./)2 !.298 9.8!2
9.22.
1?. St"%i$it t"+t:
'here was no appearance of turbidity and hence no evidence of microbial growth
when all the formulations were incubated for not less than !. days at )9RC to )?RC in case of
fluid thioglycolate medium and at 29RC to 2?RC in the case of soyabean-casein digest
medium. 'he preparations being e*amined therefore passed the test for sterility.
Fig. No. :2 St"%i$it t"+t !o% gati!$o>a#in in5+it( g"$ in +o4"an #a+"in m"1i(m
Fig. No. :2 St"%i$it t"+t !o% #,$o%am.,"ni#o$ in5+it( g"$ in t,iog$#o$at" m"1i(m
11. In )it%o "!!i#a#:
'he results of in vitro efficacy studies are shown in table )/. 'he study indicated that
Gatiflo*acin and chloramphenicol retained its antimicrobial efficacy when formulated as an in
situ gelling system and the drug was active against the selected strains of micro-organisms.
'he +one of inhibition observed for selected micro-organisms is shown in fig !2, !5, !8, 29.
Ta4$".no: 2; In )it%o "!!i#a# o! .%".a%"1 in +it( g"$+ o! gati!$o>a#in
Stan1a%1
EOI
7mm8
Fo%m($ation+ 7EOI8 mm P"%#"nt
"!!i#i"n#
F1 F2 F2 F3
4seudomonas
aeruginosa
25 25 25 25 25 !99
;taphylococcus
aureus
22 22 22 22 22 !99
Fig. No. :3 In )it%o "!!i#a# o! !o%m($ation F1 o! gati!$o>a#in
Fig. No. :: In )it%o "!!i#a# o! !o%m($ation F3 o! gati!$o>a#in
Ta4$" no: 2< In )it%o "!!i#a# o! !o%m($ation+ o! #,$o%am.,"ni#o$
Mi#%oo%gani+m+
S"$"#t"1
Stan1a%1
EOI
7mm8
Fo%m($ation+ 7EOI8 mm P"%#"nt
"!!i#i"n#
F1 F2 F2 F3
4seudomonas
#eruginosa
2/ 2/ 2/ 2/ 2/ !99
;taphylococcus
#ureus
2! 2! 2! 2! 2! !99
Fig. No. :; In )it%o "!!i#a# o! !o%m($ation F1A F2A F2A F3 o! #,$o%am.,"ni#o$.
Chapter-5 Conclusion
;. SUMMERY:
Irom the e*perimental results,
(phthalmic in situ gelling system can be formulated using pola*amer as
thermo sensitive polymer along with chitosan as viscosity enhancing agent.
'he p3 of all formulations was found to be satisfactory thus there would be
no irritation to the patient upon administration of the formulation.
'he drug content was within acceptable range which ensured dose uniformity
in the formulation. 'he drug content was ma*imum for I. of
chloramphenicol and I) of gatiflo*acin.
Gelation studies revealed that, the in situ gelling systems formed gels
instantaneously when contacted with ;'I. 'he formed gels would enhance
ocular contact time of Gatiflo*acin, chloramphenicol in eye.
Irom the rheological studies, it was concluded that, formulations e*hibited
4seudoplastic rheology. 'he viscosity was ma*imum for I. of
chloramphenicol and gatiflo*acin because of the higher concentration of
pola*amer along with chitosan. 'he ocular residence time of drug would be
prolonged because of the high viscosity.
'he in vitro release studies revealed that, drug release was slowest for I. of
gatiflo*acin, chloramphenicol after / hours.
'he results of &inetic data treatment suggested that, I! and I. followed +ero
order &inetics whereas I2 and I) followed first order &inetics.
,t was also observed that formulations I!, I) and I. followed 3iguchi
matri* which suggested diffusion controlled release.
'he bnA values obtained from 4eppas G Norsmeyer e%uation suggested that,
all the formulations showed drug release by Iic&ian diffusion mechanism.
,n vitro efficacy studies reveal that, all the formulations of gatiflo*acin and
chloramphenicol showed the !99> efficacy.
Chapter-5 Conclusion
CONCLUSION:
'he thermoreveresible insitu gelling systems of gatiflo*acin and chloramphenicol had
been prepared and evaluated. Results proved the invitro efficacy of the gatiflo*acin,
chloramphenicol formulations as well as the sustained release had also been achieved. 'he in
vitro release studies showd that chloramphenecol formulations shown better sustained release
than gatiflo*acin formulations due to low permeability of the chloramphenicol. 'a&ing this
into consideration, many poorly bioavailable drugs intended for ocular delivery can be
formulated as insitu gelling systems so can their bioavailability and efficacy are enhanced.
'he advantage that more residence time should be ta&en into account when formulating drug
with narrow therapeutic inde*. 'he thermoreversible insitu gelling system is a promising
approach for the ocular delivery of the drugs overcoming the hurdles of bioavalilability and
wash off.
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